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Orthopaedic Proceedings
Vol. 84-B, Issue SUPP_I | Pages - 12
1 Mar 2002
Gallagher J Tierney P Murray P O’Brien M
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Introduction: The infrapatellar fat pad was first described in 1904 by Albert Hoffa. Sometimes disregarded, it is apparent that the infrapatellar fat pad is of importance to knee joint function as fat at this site is only lost in severely emaciated individuals. Also, recent MRI studies have described various pathological changes affecting the fat pad. This study examined the anatomy of the infrapatellar fat pad in relation to knee symptoms and surgical approaches. Materials and Methods: 8 preserved knees were dissected via semicircular parapatellar incisions extending from the tibial tubercle to the superior patellar border and including the quadriceps muscle 13 cm above the superior border of the patella. The synovial membrane of the joint and the ligamentum mucosum were divided and the tibial tubercle was then excised. The resultant tissue complex was removed and the fat pad dissected away from surrounding structures. The appearance, volume and presence of any clefts in the pad were recorded. The cadaveric dissections were then compared to direct observation of the fat pad during total knee replacement, during arthroscopy and on MR imaging. Results: The infrapatellar fat pad was found to be present in all cases. It had a consistent shape consisting of a central mass with medial and lateral extensions. The ligamentum mucosum was attached to the intercondylar notch of the femur in all cases and measured an average of 15.7mm at its base. A horizontal cleft was found in 6 cases and a vertical cleft was found in 7 cases. Both have been previously noted. A tag extended superiorly from the posterior aspect of the fat pad in 7 cases. The volume of the fat pad had quite a large range among individual cadavers (average volume was 24 ml, range: 12–36ml). The intra-individual variation was smaller with an average difference of 4ml (range:2.7ml) between knees. Discussion: The infrapatellar fat pad has been implicated in a wide variety of conditions affecting the knee joint. It has been shown to be involved in arthofibrosis of the knee following surgery, patellar tendonitis, formation of intra-articular fibrous bands, and a site of an ossifying chondroma. It seems that fat pad pathology is usually secondary to other knee joint pathology and primary involvement is rare. The presence of clefts in the fat pad is of importance as a distended cleft may mimic an abnormality and an abnormality in the cleft may be overlooked on imaging of the knee joint. The appearance of the fat pad on direct visualisation in the living person presented a fat pad with a more globular appearance than that seen in the cadaver. The clefts were clearly visualised on MRI. Conclusion: The infrapatellar fat pad is a structure that is consistently present in the knee joint. It consists of a central body with medial and lateral and medical extensions. It usually contains a vertical cleft located superiorly and a horizontal cleft located inferiorly as well as a tag of fat extending superiorly, which forms the roof of the vertical cleft. The infrapatellar fat pad is attached to the intercondylar notch of the femur by the ligamentum mucosum and is firmly anchored to the patella by dense fibrous tissue


Orthopaedic Proceedings
Vol. 88-B, Issue SUPP_I | Pages 103 - 103
1 Mar 2006
Bohnsack M Hurschler C Wilharm A Ruehmann O Stukenborg-Colsman C Wirth C
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Purpose: The study was designed to evaluate the biomechanical and neurohistological properties of the infrapatellar fat especially concerning its potential role in the anterior knee pain syndrome. Methods: Isokinetic knee extension from 120 of flexion to full extension was simulated on 10 human knee cadaver specimens (6 male, 4 female, average age at death 44 years). Joint kinematics was evaluated by ultrasound sensors (CMS 100TM, Zebris, Isny, Germany), and retro-patellar contact pressure was measured using a thin-film resistive ink pressure system (K-ScanTM 4000, Tekscan, Boston). The infrapatellar tissue pressure was analyzed using a closed sensor cell. The patellar contact pressure was measured before and after resection of the infrapatellar fat pad. The distribution of nerve fibres in the infrapatellar fat pad was assed immunohistologically in a second part of the study. Results: Infrapatellar tissue pressure significantly increased during knee extension < 20 and flexion > 100 ranging from 343 (223) mbar at O- to 60 (64) mbar at 60 of flexion. Total resection of the infrapatellar fat pad resulted in a significant decrease in tibial external rotation of 3° in full knee extension (p=0.011), combined with a significant medial translation of the patella between 29 and 69° knee flexion (p=0.017 to 0.028). Retropatellar contact pressure was significantly (p< 0.05) reduced at all flexion angles, at 120° knee flexion more than in full knee extension. Studying all the detectable nerves present in 50 fields (x200 objective) we found an average of 6.4 substance-P- (25%) of a total of 24.7 nerve fibres in the infrapatellar fat pad. There was a significantly (p< 0.01) higher number of substance-P-fibers (24.4 (28%) of 105.7) in the superficial synovial tissue. The number of S-100-fibers was significantly (p< 0.05) higher in the central and lateral part of the fat pad. Conclusions: Based on these results, we conclude that resection of the infrapatellar fat pad could potentially reduce clinical symptoms in the anterior knee pain syndrome, and that, contrary to commonly believed, the infrapatellar fat pad may have a biomechanical function and play a role in the anterior knee pain syndrome


Orthopaedic Proceedings
Vol. 90-B, Issue SUPP_II | Pages 368 - 368
1 Jul 2008
Khan W Adesida A Andrew J Hardingham T
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Introduction: Autologous chondrocytes harvested from articular cartilage are being used for the repair of focal cartilage defects. The procedure involves injury to the cartilage and alternative sources of stem cells for use in repair are being explored. Stem cells have been found in many tissue including bone marrow and the infrapa-tellar fat pad. Infrapatellar fat pad derived stem cells present a viable and easily accessible source of stem cells for the repair of cartilage defects and tissue engineering applications. Hypoxia has been shown to improve chondrogenesis in stem cells derived from the bone marrow. We explore the hypothesis that this effect would also apply to stem cells derived from the infrapatellar fat pad. Materials and methods: Cell aggregates from early passage stem cells isolated from the infrapatellar fat pad were placed in chondrogenic media for 14 days either in a normoxic (20% oxygen) or hypoxic (5% oxygen) environment. Gene expression analysis, DNA and glycosoaminoglycan assays and immunohistochemi-cal studies were performed on the aggregates to assess chondrogenesis. Results: Cells grown under hypoxic conditions showed significantly improved chondrogenesis as determined by relatively higher gene expression of proteoglycans, collagens and SOX genes. The cell aggregates also had a higher glycosoaminoglycan content and glycosoamino-glycan content per DNA. Immunohistochemical studies confirm enhanced production of collagen types I and II and aggrecan. Discussion: These findings confirm the previously documented effects of hypoxic culture conditions on stem cells and extend the findings to include infrapatellar fat pad derived stem cells. Our findings suggest that oxygen tension has a role in regulating the function of stem cells as they undergo chondrogenesis. In culture these cells appear to function optimally in an atmosphere of reduced oxygen that more closely approximates documented in vivo oxygen tension. This has important implications in future tissue engineering applications of these cells


Orthopaedic Proceedings
Vol. 86-B, Issue SUPP_III | Pages 372 - 372
1 Mar 2004
Bohnsack M Meier F Schmolke S Walter G Wirth C RŸhmann O
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Aims: The purpose of the study was to determine the distribution and speciþcation of nerve þbers in the infrapatellar fat pad especially concerning nociceptive substance-P þbres. Methods: The infrapatellar fat pad was taken as a fresh specimen out of 21 patients (4 male, 17 female, mean age 69 years) during total knee arthroplasty. It was dissected in þve deþned parts, þxed and embedded in parafþn. Immunohistochemical techniques using antibodies against S-100 protein and substance-P were employed to determine and specify the nerve þbres. Results: Studying all the detectable nerves present in 50 þelds (x200 objective) we found an average of 6,4 substance-P- (25%) of a total of 24,7 nerve þbres in the infrapatellar fat pad. There was a significantly (p< 0,01) higher number of substance-P-þbers (24,4 (28%) of 105,7) in the surfacing synovial tissue. The number of S-100-þbers was signiþcantly (p< 0,05) higher in the central and lateral part of the fat pad. Conclusions: The occurance and distribution of nerve þbres in the infrapatellar fat pad suggests a nociceptive function. A neurohistological role in the anterior knee pain syndrome is assumed


Orthopaedic Proceedings
Vol. 86-B, Issue SUPP_III | Pages 254 - 254
1 Mar 2004
Bohnsack M Wilharm A Demirtas T Rühmann O Wirth C Hurschler C
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Aims: This biomechanical study was performed to evaluate the consequences of a total infrapatellar fat pad resection on knee kinematics and patellar contact pressure. Methods: Knee motion between 120∞ of flexion and full extension was performed in a knee kinemator on 10 fresh frozen knee specimens (6 male, 4 female, average age 44 years). The joint kinematics was evaluated by ultrasound sensors (Zebris-system), the patellar contact pressure was measured using a thin-film resistive ink pressure system (Tekscan). All data were taken before and after resection of the infrapatellar fat pad and statistically analyzed. Results: A total resection of the infrapatellar fat pad resulted in a significant (p< 0,05) decrease of the tibial external rotation in knee extension combined with a significant (p< 0,05) medial translation of the patella. The patellar contact pressure was significantly (p< 0,05) reduced, in knee flexion more than in knee extension. Conclusions: We conclude that a resection of the infrapatellar fat pad might reduce clinical symptoms in the anterior knee pain syndrome. A biomechanical function of the infrapatellar fat is suspected


Orthopaedic Proceedings
Vol. 91-B, Issue SUPP_III | Pages 424 - 424
1 Sep 2009
Khan WS Anand S Tew S Andrew JG Johnson DS Hardingham TE
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There is an ever-increasing clinical need for the regeneration and replacement of tissue to replace soft tissue lost due to trauma, disease and cosmetic surgery. A potential alternative to the current treatment modalities is the use of tissue engineering applications using mesenchymal stem cells that have been identified in many tissue including the infrapatellar fat pad. In this study, stem cells isolated from the infrapatellar fat pad were characterised to ascertain their origin, and allowed to undergo adipogenic differentiation to confirm multilineage differentiation potential. The infrapatellar fat pad was obtained from total knee replacement for osteoarthritis. Cells were isolated and expanded in monolayer culture. Cells at passage 2 stained strongly for CD13, CD29, CD44, CD90 and CD105 (mesenchymal stem cell markers). The cells stained poorly for LNGFR and STRO1 (markers for freshly isolated bone marrow derived stem cells), and sparsely for 3G5 (pericyte marker). Staining for CD34 (haematopoetic marker) and CD56 (neural and myogenic lineage marker) was negative. For adipogenic differentiation, cells were cultured in adipogenic inducing medium consisting of basic medium with 10ug/ml insulin, 1uM dexamthasone, 100uM indomethacin and 500uM 3-isobutyl-1-methyl xanthine. By day 16, many cells had lipid vacuoles occupying most of the cytoplasm. On gene expression analyses, the cells cultured under adipogenic conditions had almost a 1,000 fold increase in expression of peroxisome proliferator-activated receptor gamma-2 (PPAR gamma-2) and 1,000,000 fold increase in expression of lipoprotein lipase (LPL). Oil red O staining confirmed the adipogenic nature of the observed vacuoles and showed failure of staining in control cells. Our results show that the human infrapatellar fat pad is a viable potential autogeneic source for mesenchymal stem cells capable of adipogenic differentiation as well as previously documented ostegenic and chondrogenic differentiation. This cell source has potential use in tissue engineering applications


Orthopaedic Proceedings
Vol. 92-B, Issue SUPP_I | Pages 59 - 59
1 Mar 2010
Khan* W Anand S Tew S Johnson D Andrew J Hardingham T
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There is an ever-increasing clinical need for the regeneration and replacement of tissue to replace soft tissue lost due to trauma, disease and cosmetic surgery. A potential alternative to the current treatment modalities is the use of tissue engineering applications using mesenchymal stem cells that have been identified in many tissue including the infrapatellar fat pad. In this study, stem cells isolated from the infrapatellar fat pad were characterised to ascertain their origin, and allowed to undergo adipogenic differentiation to confirm multilineage differentiation potential. The infrapatellar fat pad was obtained from total knee replacement for osteoarthritis. Cells were isolated and expanded in monolayer culture. Cells at passage 2 stained strongly for CD13, CD29, CD44, CD90 and CD105 (mesenchymal stem cell markers). The cells stained poorly for LNGFR and STRO1 (markers for freshly isolated bone marrow derived stem cells), and sparsely for 3G5 (pericyte marker). Staining for CD34 (haematopoetic marker) and CD56 (neural and myogenic lineage marker) was negative. {BR}For adipogenic differentiation, cells were cultured in adipogenic inducing medium consisting of basic medium with 10ug/ml insulin, 1uM dexamthasone, 100uM indomethacin and 500uM 3-isobutyl-1-methyl xanthine. By day 16, many cells had lipid vacuoles occupying most of the cytoplasm. On gene expression analyses, the cells cultured under adipogenic conditions had almost a 1,000 fold increase in expression of peroxisome proliferator-activated receptor gamma-2 (PPAR gamma-2) and 1,000,000 fold increase in expression of lipoprotein lipase (LPL). Oil red O staining confirmed the adipogenic nature of the observed vacuoles and showed failure of staining in control cells. Our results show that the human infrapatellar fat pad is a viable potential autogeneic source for mesenchymal stem cells capable of adipogenic differentiation as well as previously documented ostegenic and chondrogenic differentiation. This cell source has potential use in tissue engineering applications


Orthopaedic Proceedings
Vol. 92-B, Issue SUPP_I | Pages 67 - 68
1 Mar 2010
Chandrashekar S Arumilli B Conway A Hinduja K Paul A Freemont A
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Introduction: Anterior knee pain is a very common presenting symptom. Fat pad syndrome is an uncommon and a difficult condition to manage. The diagnosis is usually reached after a period of physiotherapy and investigation to rule out the more common aetiologies of anterior knee pain. Patients & Methods: All patients who underwent excision of the infrapatellar fat pad following a diagnosis of Fat pad syndrome are included. Each patient was evaluated to exclude patellofemoral problems and intraarticular pathologies as the cause of anterior knee pain. Each patient underwent MR imaging and all the excised specimens were sent for histological analysis. Results: The MR imaging provided with the provisional diagnosis in all patients. All the specimens were examined by a single senior histopathologist to correlate with the provisional diagnosis. The histology confirmed Hoffa’s syndrome in 5 patients and in the remaining 15 patients a spectrum of rare diagnoses as suspected by Magnetic Resonance imaging. The more notable conditions were two synovial sarcomas, three haemangiomas and a Giant cell tumour of the tendon sheath. All patients were treated successfully with complete excision. No recurrences were recorded at the end of 3 year follow-up and all patients were symptom free. Conclusion: The work up of a patient with suspected infrapatellar fat pad syndrome must include MR imaging and the exact underlying pathology should be confirmed with histological analysis of the excised fat pad as the rare causes include soft tissue malignancy


Orthopaedic Proceedings
Vol. 102-B, Issue SUPP_2 | Pages 31 - 31
1 Feb 2020
Acuña A Samuel L Yao B Faour M Sultan A Kamath A Mont M
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Introduction. With an ongoing increase in total knee arthroplasty (TKA) procedural volume, there is an increased demand to improve surgical techniques to achieve ideal outcomes. Considerations of how to improve post-operative outcomes have included preservation of the infrapatellar fat pad (IPFP). Although this structure is commonly resected during TKA procedures, there is inconsistency in the literature and among surgeons regarding whether resection or preservation of the IPFP should be achieved. Additionally, information about how surgical handling of the IPFP influences outcomes is variable. Therefore, the purpose of this systematic review was to evaluate the influence of IPFP resection and preservation on post-operative flexion, pain, Insall-Salvati Ratio (ISR), Knee Society Score (KSS), patellar tendon length (PTL), and satisfaction in primary TKA. Methods. A systematic literature search was performed to retrieve all reports that evaluated IPFP resection or preservation during total knee arthroplasty (TKA). The following databases were queried: PubMed, EBSCO host, and SCOPUS, resulting in 488 unique reports. Two reviewers independently reviewed the studies for eligibility based on pre-established inclusion and exclusion criteria. A total of 11 studies were identified for final analysis. Patient demographics, type of surgical intervention, follow-up duration, and clinical outcome measures were collected and further analyzed. This systematic review reported on 11,996 total cases. Complete resection was implemented in 3,723 cases (31%), partial resection in 5,458 cases (45.5%), and preservation of the IPFP occurred in 2,815 cases (23.5%). Clinical outcome measures included patellar tendon length (PTL) (5 studies), knee flexion (4 studies), pain (6 studies), Knee Society Score (KSS) (3 studies), Insall-Salvati Ratio (ISR) (3 studies), and patient satisfaction (1 study). Results. There were no differences found following IPFP resection for patient satisfaction (p=0.92), ISR (all p-values >0.05), and KSS (all p-values >0.05). Mixed evidence was found for patellar tendon length, pain, and knee flexion following IPFP resection vs. preservation. Conclusion. Given the current literature and available data, there were several clinical outcome measures that indicated better patient results with preservation of IPFP during primary TKA in comparison to the resection of IPFP. Specifically, resection resulted in inferior outcomes for patellar tendon length, knee flexion, and pain measurements. However, more extensive research is needed to better determine that preservation is the superior surgical decision. This includes a need for more randomized controlled trials (RCTs). Future studies should focus on conditions in which preservation or resection of IPFP would be best indicated during TKA in order to establish guidelines for best surgical outcomes in those patients. For any figures or tables, please contact authors directly


Orthopaedic Proceedings
Vol. 90-B, Issue SUPP_II | Pages 324 - 324
1 Jul 2008
Khan W Johnson D Andrew J Hardingham T
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Introduction: In this study infrapatellar fat pad (IPFP) derived stem cells were expanded with and without Fibroblast Growth Factor-2 (FGF-2) supplementation and were compared with regards to their ability to proliferate and differentiate into chondrocytes. Materials and Methods: Cells were isolated from the IPFP tissue and expanded in monolayer culture with and without rhFGF-2 supplementation (final concentration 10ng/ ml). Cell aggregates were placed in chondrogenic media for two weeks. Gene expression studies were carried out using quantitative real time PCR. Immunohistochemical labelling was performed with antibody localisation determined by an immunoperoxidase procedure. The pellets were also weighed and digested in papain for DNA and glycosoaminoglycan (GAG) analysis. Results: Cells expanded in FGF-2 supplemented media were smaller and proliferated more rapidly. The FGF-2 supplemented cell aggregates also showed 100 times higher expression of collagen type II (COL2A1). Immunohistochemical studies showed that pellets made from FGF-2 treated cells stained more strongly for collagen II and more weakly for collagen I. Pellets made with FGF-2 treated cells were larger, continued with enhanced proliferation and contained more proteoglycan. Conclusion: Our findings show enhanced proliferation and chondrogenic differentiation in IPFP derived stem cells expanded in FGF-2 supplemented media


Orthopaedic Proceedings
Vol. 94-B, Issue SUPP_XXXIX | Pages 215 - 215
1 Sep 2012
Eireamhoin S Buckley C Schepens A Jones E McGonagle D Mulhall K Kelly D
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Although chondrocytes have been used for autologous implantation in defects of articular cartilage, limited availability and donor-site morbidity have led to the search for alternative cell sources. Mesenchymal stem cells from various sources represent one option. The infrapatellar fat-pad is a promising source. Advantages include low morbidity, ease of harvest and ex-vivo evidence of chondrogenesis. Expansion of MSCs from human fat-pad in FGF-2 has been shown to enhance chondrogenesis. To further elucidate this process, we assessed the role of TGF-?3, FGF-2 and oxygen tension on growth kinetics of these cells during expansion.

Methods

Infrapatellar fatpads were obtained from 4 donors with osteoarthritis. Cells were expanded in various media formulations (STD, FGF, TGF and FGF/TGF) at both 20% and 5% oxygen tensions. Colony forming unit fibroblast assays were performed for each expansion group and assessed with crystal violet staining. Cell aggregates from each group underwent chondrogenic differentiation in 5% and atmospheric oxygen tension. Pellets were analyzed on day 21.

Results

5% Oxygen tension during expansion increased the colony size for both FGF and FGF/TGF groups. Cells expanded in FGF/TGF proliferated more rapidly. Biochemical analysis revealed that cells expanded in FGF-2 had higher glycosaminoglycan synthesis rates, a marker for chondrogenesis. Differentiation at 5% pO2 led to higher levels of sGAG but its effect was generally less potent compared to expansion in FGF-2.


Orthopaedic Proceedings
Vol. 92-B, Issue SUPP_I | Pages 161 - 161
1 Mar 2010
Seo S Kim C Ha D Chung H
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The purpose of this study in to investigate the role of infrapatellar fat pad on primary total knee arthroplasty. We evaluated 100 patients who had been undergone TKA from August 2002 to July 2003, with open box posterior substituting femoral component implant (Scorpio PS Knee. ™. ). The study was performed prospectively and randomly allocated. We divided two groups. Group 1 (50 knees) was preserved infrapatellar fat pad and repaired fad at wound closure. Group 2 (50 knees) was excised infrapatellar fat pad as possible and repaired only joint capsule. We analyzed and compared clinical results of Knee Society knee (KS) score, function score, patellar score and Insall-Salvati ratio in both groups. The complications of each group were evaluated. Patients were followed up for mean 40 months(17~52 months). Mean KS score was 91.9 (91.94±5.58) in Group 1 and 90.9(90.92±6.38) in Group 2. Mean function score was 81.6(81.64±13.18) in Group 1 and 83.7(83.79±17.71) in Group 2. Mean patellar score was 29.9(29.89±9.10) in Group 1 and 27.9(27.90±1.80) in Group 2. And mean patellar height as Insall-Salvati ratio was 1.19(1.19±0.17) in Group 1 and 1.23(1.23±0.11) in Group 2. The differences between the Group 1 and Group 2 in all of index were statistically insignificant. In complications, 2 cases of recurrent hemarthrosis were observed in Group 1 patients. We concluded The difference of clinical outcomes whether infrapatellar fat pad was excised or not were statistically insignificant. However, preservation of infrapatellar fat pad on open boxed PS TKA showed unique complications such as recurrent hemarthrosis which might be caused by fat pad adhesion to intercondylar notch. We propose that infrapatellar fat pad on primary PS TKA with open box design would like to be excised for prevention of unique complications


Orthopaedic Proceedings
Vol. 102-B, Issue SUPP_11 | Pages 122 - 122
1 Dec 2020
Huri PY Talak E Kaya B Huri G
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Articular cartilage is often damaged, and its treatment is usually performed by surgical operation. Today, tissue engineering offers an alternative treatment option for injuries or diseases with increasing importance. Infrapatellar fat pad (IPFP) is a densely vascularized and innervated extra synovial tissue that fills the anterior knee compartment. Adipose-derived stem cells from infrapatellar fat pad (IPFP-ASCs) have multipotency means that they can differentiate into connective tissue cells and have age-independent differentiation capacity as compared to other stem cells. In this study, the osteochondral tissue construct was designed with different inner pattern due to original osteochondral tissue structure and fabrication of it was carried out by 3D printing. For this purpose, alginate (3% w/v) and carboxymethylcellulose (CMC) (9%w /v) were used as bioink. Also, IPFP-ASCs were isolated with enzymatic degradation. Osteogenic and chondrogenic differentiation of IPFP-ASCs were investigated with Alizarin Red and Alcian Blue staining, respectively. IPFP-ASCs-laden osteochondral graft differentiation will be induced by controlled release of growth factor BMP-2 and TGF-β. Before this step, nanocapsules formation with double emission technique with model protein BSA was carried out with different concentration of PCL (5%,10% and 20%). The morphology and structure of the nanocapsules were determined with scanning electron microscopy (SEM). Also, we successfully designed and printed alginate and CMC based scaffold with 20 layers. Chondrogenic and osteogenic differentiation of IPFP-ASCs with suitable culture conditions was obtained. The isolation of IPFP-ASCs, formation of the nanocapsules, and 3D printing of osteochondral graft were carried out successfully


Orthopaedic Proceedings
Vol. 94-B, Issue SUPP_XXXVI | Pages 50 - 50
1 Aug 2012
Hopper N Wardale J Rushton N
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Introduction. Mesenchymal stem cells (MSC) are an attractive cell population for regeneration of mesenchymal tissue such as bone and cartilage. Various studies have demonstrated the repair capacity of MSCs and even their usefulness in treating critical size defects. Much of the work conducted on adult stem cells has focused on MSCs found within the bone marrow stroma. Adipose tissue, like bone marrow, is derived from the embryonic mesenchyme and contains a stroma that is easily isolated. The aim of the present study is to evaluate the differentiation capability of adipose-tissue derived stem cells (ASC) extracted from the infrapatellar fat pad. Materials and Methods. Human infrapatellar fat pad tissue was obtained from patients undergoing total joint replacement for osteoarthritis with full ethical consent. A multipotent progenitor cell population was derived after collagenase digestion from the adipose tissue. The ASCs were induced to differentiate towards adipogenic, chondrogenic, and osteogenic lineages for 21 days both in normoxic and hypoxic cell culture conditions. The differentiation and multilineage potential was assessed according to cell morphology and in vitro detection of tissue-specific differentiation molecules. Results. After 3 weeks in culture the staining for oil-red-o, alcian bue, and alizarin-red confirmed the differentiation capability of ASC's to adipogenic, chondrogenic, and osteogenic lineages, respectively. The hypoxic cell culture condition was found to support the ASCs' chondrogenic differentiation capability and subsequently enhanced the proteoglycan release from the cells. Fluorescence-activated cell sorting (FACS) confirmed the presence of stromal precursor cell marker STRO-1 in the ASC population. Discussion. Subcutaneous adipose tissue is particularly attractive reservoir for progenitor cells because it is easily accessible, rather abundant, and self-replenishing. The results of this study demonstrate that ASCs can be derived from infrapatellar fat pad and that they have potential for musculoskeletal tissue repair and regeneration. Further studies are underway to evaluate how to adopt a biomaterial to deliver these cells into the defect area to facilitate the healing response


Orthopaedic Proceedings
Vol. 92-B, Issue SUPP_I | Pages 161 - 161
1 Mar 2010
Itokawa T Kondo M Tsumura H Fujii T Azuma T Tomari K Kadoya Y
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Achieving deep flexion of knee after total knee arthroplasty (TKA) is particularly desirable in some Asian and Middle Eastern who have daily or religious customs typically use full knee flexion. After TKA, some patients complained about anterior knee pain during deep knee flexion. We evaluated the efficacy of arthroscopic fat pad resection in a series of patients suffering from anterior knee pain associated with high flexion achievement after TKA. The efficacy of fat pad resection via arthroscopy for treating anterior knee pain associated with high flexion angle (average = 133.1°) was evaluated in eight knees of eight patients among 207 knees performed between 1996 and 1999. The mean age of patients was 71.1 years when the primary TKA was performed. All implatants were posterior stabilized type (IB-II, Nexgen PS and LPS). The symptom of anterior knee pain during deep knee flexion developed within one year after TKA in all cases. In addition to pain in eight knees, two patients have crepitation as the knee was flexed and extended and three patients had hydrarthrosis. Impingement and fibrosis of fat pad were confirmed, and fibrous structures were removed by arthroscopy. Before arthroscopy, the symptom obviously subsided after injection of local anesthesia into infrapatellar fat pad. Patellar clunk syndrome is also soft tissue impingement and suprapatellar fibrous nodule becomes entrapped intercondylar notch on the femoral component during knee flexion. On this point, these cases does not cause by patellar clunk syndrome. After fat pad resection, the symptom disappeared, and keeps symptom-free after a mean follow-up of six years five months in all cases. Any complications following fat pad resection, such as patella baja and necrosis, were not experienced. Those cases achieving higher flexion angle tended to experience severe pain and shorter time interval between TKA and arthroscopic surgery, suggesting impingement of the infrapatellar fat pad is closely related to deep flexion after TKA. These results demonstrate that the anterior knee pain due to repetitive infrapatellar fat pad impingement is one of the complications during deep knee flexion after TKA, and the arthroscopic fat pad resection is useful to relief the anterior knee pain. Because of our experience with patients encountering anterior knee pain, we have begun to remove 70 to 80% of the fat pad during the primary TKA procedure since 1999, and until today, none developed anterior knee pain thought to be associated with fat pad impingement, patellar baja nor patellar necrosis. We suggest that fat pad resection is necessary to prevent the anterior knee pain due to fat pad impingement during deep flexion in TKA


Orthopaedic Proceedings
Vol. 102-B, Issue SUPP_11 | Pages 30 - 30
1 Dec 2020
Haartmans M Eveque-Mourroux M Eijkel G Emanuel K Tuijthof G van Rhijn L Heeren R Emans P Cillero-Pastor B
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The incidence of osteoarthritis (OA) is increasing in our younger population. OA development early in life is often related to cartilage damage, caused by (sport) injury or trauma. Detection of early knee OA is therefore crucial to target early treatment. However, early markers for OA prognosis or diagnosis are lacking. Hoffa's fat pad (HFP) is an emerging source for knee biomarkers, as it is easily accessible and shows important interaction with the homeostasis of the knee. In this study, we used Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) as a first approach. MALDI-MSI allows the study of tissue-specific molecular distributions. Therefore, we used MALDI-MSI to analyze the lipid profiles in the HFP of three patients with OA and three patients undergoing cartilage regenerative treatment. We demonstrate that the lipid profile of patients with OA is different from patients with cartilage defects.

HFP of each patient were snap frozen directly after surgical resection and cryosectioned at 15 μm. Each slide was sublimed with Norharmane matrix and analyzed by MALDI-MSI in positive and negative ion modes at a lateral resolution of 50 μm on a RapifleX Tissue Typer. The difference between patient groups were analyzed using principle component analysis and linear discriminant analysis. Lipid identifications were obtained on an Orbitrap Elite™ Hybrid Ion Trap-Orbitrap Mass Spectrometer in data dependent acquisition mode and analyzed using Lipostar software.

Linear discriminant analysis showed a specific lipid profile for each group (variance 33.94%). Score projections revealed a differential lipid spatial distribution of OA patients compared to cartilage defect patients. Among the lipids that differed significantly, for instance, the m/z 760.59 [M+H]+ was associated to osteoarthritis and identified as glycerophospholipid (PC 34:1), a main component of biological membranes. Additionally, the samples were found to be intra-tissue heterogeneous, with molecular profiles found in adipose-, connective- and synovial tissue.

These results suggest that lipid profiles in HFP could be useful for early OA detection. However, intra-tissue heterogeneity in HFP should be recognized when using HFP as a biomarker source.


Orthopaedic Proceedings
Vol. 105-B, Issue SUPP_2 | Pages 52 - 52
10 Feb 2023
Di Bella C
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3D printing and Bioprinting technologies are becoming increasingly popular in surgery to provide a solution for the regeneration of healthy tissues. The aim of our project is the regeneration of articular cartilage via bioprinting means, to manage isolated chondral defects. Chrondrogenic hydrogel (chondrogel: GelMa + TGF-b3 and BMP6) was prepared and sterilised in our lab following our standard protocols. Human adipose-derived mesenchymal stem cells were harvested from the infrapatellar fat pad of patients undergoing total knee joint replacements and incorporated in the hydrogel according to our published protocols. The chondrogenic properties of the chondrogel have been tested (histology, immunohistochemistry, PCR, immunofluorescence, gene analysis and 2. nd. harmonic generation microscopy) in vitro and in an ex-vivo model of human articular defect and compared with standard culture systems where the growth factors are added to the media at repeated intervals. The in-vitro analysis showed that the formation of hyaline cartilage pellet was comparable between the two strategies, with a similar metabolic activity of the cells. These results have been confirmed in the ex-vivo model: hyaline-like cartilage was observed within the chondral defect in both the chondrogel group and the control group after 28 days in culture. The use of bioprinting techniques in vivo requires the ability of stem cells to access growth factors directly in the environment they are in, as opposed to in vitro techniques where these factors are provided externally at recurrent intervals. This study showed the successful strategy of incorporating chondrogenic growth factors for the formation of hyaline-like cartilage in vitro and in an ex-vivo model of chondral loss. The incorporation of chondrogenic growth factors in a hydrogel is a possible strategy for articular cartilage regeneration


Orthopaedic Proceedings
Vol. 98-B, Issue SUPP_3 | Pages 152 - 152
1 Jan 2016
Sekiya H Takatoku K Takada H Kanaya Y Sasanuma H
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From October 2005 to March 2014, we performed 46 arthroscopic surgeries for painful knee after knee arthroplasty. We excluded 16 cases for this study such as, unicompartmental knee arthroplasty, infection, patellar clunk syndrome, patellofemoral synovial hyperplasia, aseptic loosening, and follow-up period after arthroscopic surgery less than 6 months. Thirty cases matched the criteria. They had knee pain longer than 6 months after initial total knee arthroplasty (TKA), they had marked tenderness at medial and/or lateral tibiofemoral joint space, and also they complained walking pain with or without resting pain. Twenty one cases had initial TKA at our institute. In consideration of total number of TKA (n=489) in the period at our institute, incident rate of painful knee after initial TKA was 4.3%. Of 30 cases, 3 cases were male, and 27 cases were female. Types of implant were 4 in cruciate retaining type, 1 in cruciate substituting type, and 25 in posterior stabilized type. Age at the arthroscopy was 72 years old (51–87 years old), and period form initial TKA to pain perception was 18 months(1 – 144 months), and period from initial TKA to arthroscopic surgery was 29 months (6 – 125 months), and follow-up period after arthroscopy was 36 months (6 – 93 months). All arthroscopic debridement were performed through 3 portals, anteromedial, anterolateral, and proximal superomedial portal. Scar tissue impingements more than 5 mm wide were found in 87% of the cases both medial and lateral femorotibial joint spaces. Infrapatellar fat pad were covered with whitish scar tissue in all cases, and the scar tissue were connecting with the scar tissue which found at medial or lateral femorotibial joint spaces. We removed all scar tissue with motorized shaver or punches. At final follow-up, complete pain free in 63%, marked improvement in 3%, half improvement in 20%, slight improvement in 3%, and no change in 10% of the cases. Previously in the literatures, two reasons of the pain after total knee arthroplasty had been reported, patellar clunk syndrome, and patellar synovial hyperplasia. All cases reported this study had marked tenderness at tibiofemoral joint space. It was difficult to explain the tenderness by previously reported pathological mechanisms. We had to find another pathological mechanism to explain the pain of our cases. Painful knee due to scar tissue formation known as “infrapatellar contracture syndrome” after anterior cruciate ligament reconstruction surgery was previously reported. We hypothesized similar scar tissue formation should occur after TKA that caused painful knee. Continuity of the solid scar tissue between infrapatellar fat pad with the scar tissue at tibiofemoral joint space should be the cause of impingement at femorotibial joint even small size of scar tissue. From this study, we have to recognize that painful knee after TKA is not infrequent complication. And, if we could deny infection, and aseptic loosening in painful knee after TKA, arthroscopic debridement was good option to solve the pain. We could expect improvement of the pain more than half in 87% of cases


Orthopaedic Proceedings
Vol. 103-B, Issue SUPP_16 | Pages 30 - 30
1 Dec 2021
Vogt A Darlington I Brooks R Birch M McCaskie A Khan W
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Abstract. Osteoarthritis is a common articular cartilage disorder and causes a significant global disease burden. Articular cartilage has a limited capacity of repair and there is increasing interest in the use of cell-based therapies to facilitate repair including the use of Mesenchymal Stromal Cells (MSCs). There is some evidence in the literature that suggests that advancing age is associated with declining MSC function, including reduced proliferation and differentiation potential, and greater cellular apoptosis. In our study, we first performed a systematic review of the literature to determine the effects of chronological age on the in vitro properties of MSCs, and then performed a laboratory study to investigate these properties. We initially conducted a PRISMA systematic review of the literature to review the evidence base for the effects of chronological age on the in vitro properties of MSCs including cell numbers, expansion, cell surface characterization and differentiation potential. This was followed by laboratory based experiments to assess these properties. Tissue from patients undergoing total knee replacement surgery was used to isolate MSCs from the infrapatellar fat pad using a method developed in our laboratory. The growth kinetics was determined by calculating the population doublings per day. Following expansion in culture, MSCs at P2 were characterised for a panel of cell surface markers using flow cytometry. The cells were positive for CD73, CD90 and CD105, and negative for CD34 and CD45. The differentiation potential of the MSCs was assessed through tri-lineage differentiation assays. Chronological age-related changes in MSC function have important implications on the use of these cells in clinical applications for an ageing population. The results from this study will be used to plan further work looking at the effects of chronological age on cellular senescence and identify pathways that could be targeted to potentially reverse any age-related changes


Orthopaedic Proceedings
Vol. 103-B, Issue SUPP_16 | Pages 14 - 14
1 Dec 2021
Darlington I Vogt A Williams EC Brooks R Birch M Mohorianu I Khan W McCaskie A
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Abstract. Focal articular cartilage defects do not heal and, left untreated, progress to more widespread degenerative changes. A promising new approach for the repair of articular cartilage defects is the application of cell-based regenerative therapies using mesenchymal stromal cells (MSCs). MSCs are however present in a number of tissues and studies suggest that they vary in their proliferation, cell surface characterisation and differentiation. As the phenotypic properties of MSCs vary depending on tissue source, a systematic comparison of the transcriptomic signature would allow a better understanding of these differences between tissues, and allow the identification of markers specific to a MSC source that is best suited for clinical application. Tissue was used from patients undergoing total knee replacement surgery for osteoarthritis following ethical approval and informed consent. MSCs were isolated from bone, cartilage, synovium and infrapatellar fat pad. MSC number and expansion were quantified. Following expansion in culture, MSCs were characterised using flow cytometry with several cell surface markers; the cells from all sources were positive for CD44, CD90 and CD105. Their differentiation potential was assessed through tri-lineage differentiation assays. In addition, bulk mRNA-sequencing was used to determine the transcriptomic signatures. Differentially expressed (DE) genes were predicted. An enrichment analysis focused on the DE genes, against GO and pathway databases (KEGG and Reactome) was performed; protein-protein interaction networks were also inferred (Metascape, Reactome, Cytoscape). Optimal sourcing of MSCs will amplify their cartilage regeneration potential. This is imperative for assessing future therapeutic transplantation to maximise the chance of successful cartilage repair. A better understanding of differences in MSCs from various sources has implications beyond cartilage repair