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Orthopaedic Proceedings
Vol. 93-B, Issue SUPP_II | Pages 158 - 158
1 May 2011
Moretti B Notarnicola A Moretti L Patella S Setti S Ciullo F Patella V
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The employment of biophysical therapy to accelerate the healing of tissues is by now a well-established practice in many orthopaedic situations, indicated mainly for osteogenesis and chondrogenesis. Assessments of the effects of biophysical stimuli on joint cartilage (CRES, Cartilage Repair & Elecromagnetic Stimulation) performed with pre-clinical studies and clinical studies (in operations to reconstruct LCA and microfractures) have shown how biophysical stimulation controls the microambience, and have suuplied the rationale for passing to an evaluation of the effects also in the case of joint replacement.

We launched a randomized prospective clinical study of 30 patients aged between 60 and 85 years, afflicted with gonarthrosis and undergoing operation for prosthesis. The randomization involved subdividing them into two homogeneous groups: the first with biophysical treatment with I-ONE therapy (Igea-Clinical Biophysics) (experimental group); the second not undergoing the biophysical treatment (control group). In the experimenal group, the I-ONE therapy was commenced at 3–7 days from the operation, administered for 4 hours per day and maintained for 60 days consecutively. The clinical evaluations were performed by compiling functional reports (swelling of the knee, Knee Score, SF-36 and VAS) in the pre-operative period and postoperatively at 1, 2, 6 and 12 months. The data processing was subjected to statistical evaluation by an independent observer using Student’s two-tail t test and the Generalized Linear Mixed Effects Model.

The preliminary results showed that at the baseline there are no differences between the groups either for the KNEE score, nor the VAS, or the SF-36. Already after 1 month the differences between the groups are statistically significant (p< 0.05 for KNEE score, p< 0.001 for swelling, p< 0.0001 for VAS and SF-36). At 2 months the differences between the groups are highly significant (p< 0.0001). The study entails a long-term evaluation with monitoring of the patients at one year from operation.

The results of this study supply the basis for clinical employment of biophysical treatment with I-ONE immediately following joint surgery, enabling inflammation to be controlled and increasing anabolic activity and protecting its microambience.


Orthopaedic Proceedings
Vol. 91-B, Issue SUPP_III | Pages 460 - 460
1 Sep 2009
Moretti B Notarnicola A Iannone F Moretti L Garofalo R Patella V
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The purpose of this study was to investigate the effects of extra corporeal shock waves (ESW) therapy on the metabolism of healthy and osteoarthritic human chondrocytes, and particularly on the expression of IL-10, TNF-α and β1 integrin.

Human adult articular cartilage was obtained from 9 patients (6 male and 3 females), with primary knee osteoarthritis (OA), undergoing total joint replacement and from 3 young healthy donors (HD) (2 males, 1 female) with joint traumatic fracture. After isolation, chondrocytes underwent ESW treatment (Electromagnetic Generator System, Minilith SL1, Storz Medical) at different parameters of impulses, energy levels and energy fluxes. After that, chondrocytes were cultured in 24-well plate in DMEM supplemented with 10% FCS for 48 hours and then β1 integrin surface expression and intracellular IL-10 and TNF-α levels were evaluated by flow-cytometry.

At baseline, osteoarthritic chondrocytes expressed significantly lower levels of β1 integrin and higher levels and IL-10 and TNF-α levels. It has been recently reported that ESW may be useful to treat OA in dogs, and veterinarians have begun to use ESW also to treat OA in horses.

Following ESW application, while β1 integrin expression remain unchanged, a significant decrease of IL-10 and TNF-α intracellular levels was observed both in osteoarthritic and healthy chondrocytes. IL-10 levels decreased at any impulses and energy levels, while a significant reduction of TNF-α was mainly found at middle energies.

Our study confirmed that osteoarthritic chondrocytes express low β1 integrin and high TNF-α and IL-10 levels. Nonetheless, ESW treatment application down-regulate the intracellular levels of TNF-α and IL-10 by chondrocytes, suggesting that ESW might restore TNF-α and IL-10 production by osteoarthritic chondrocytes at normal levels thus potentially interfering with the pathologic mechanisms causing cartilage damage in OA and representing the theoretical rationale for using ESW as therapy of OA.


Orthopaedic Proceedings
Vol. 91-B, Issue SUPP_III | Pages 448 - 449
1 Sep 2009
Moretti B Notarnicola A Tamma R Zallone A Patella S Patella V
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The osteoclastogenesis is regulated by a complex signaling system between the pro-apoptotic factors (Bax-Cyclin E2-Cdk2) and the tumor necrosis factor family (RANKL-RANK-OPG).

Extracorporeal Shock Waves Therapy (ESWT) have recently been used in orthopaedic treatments to induce bone repair, but their mechanisms of action are not sufficiently investigated. So we studied the effect of shock-waves on murine osteoblastic cells.

Osteoblast cultures were subjected to a single shock-wave with combinations of low energy intensities (0.05mJ/mm2) and 500 number of shocks (impulses), whereas control cells received no treatment. We valued the cell viability quantifying the expressions of Bax and Opg by PCR.

We found an immediate negative effect on cell viability, that occurs with an increase of Bax protein expression after 3 hours of treatment. After a longer time lapse a stimulatory effect on cell proliferation, as reflected by the increase of a G(1)-S phase marker, was observed. In fact, in the following 24, 48 and 72 hours after ESW treatment, we found a stronger association of Cyclin E2 and Cdk2, forming active cyclin E-Cdk2 kinase, compared to untreated cells at the same times.

We further explored the molecular mechanism for the ESW induction of osteogenesis: by Real Time PCR an enhancement of Runx2 mRNA, evident 48 hours after the treatment, was found. A link between physical ESW and Runx2 activation has been already demonstrated. ESW-induced

O2- production, followed by tyrosine kinase mediated ERK activation and Runx2 activation, resulted in osteogenic cell growth and maturation. Moreover, we analyzed the cytokines RANK-L and OPG osteoblast expression, involved in regulation of osteoclastogenesis. A decrease in RANK-L/OPG ratio was found, perhaps leading to a reduced osteoclastogenesis.

The Shock waves have a repair action on bone and it can been explained by the regulation on osteoclastogenesis by the apoptoic pathway of BAX and OPG.


Orthopaedic Proceedings
Vol. 91-B, Issue SUPP_II | Pages 272 - 272
1 May 2009
Moretti B Notarnicola A Foscarini P Tamma R Zallone A Patella V
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Aims: Extracorporeal Shock Waves Therapy (ESWT) has recently been used in orthopaedic treatments to induce bone repair, but their mechanisms of action are not sufficiently investigated. So we studied the effect of shockwaves on murine osteoblastic cells.

Methods: Osteoblast cultures were subjected to a single shockwave with combinations of low energy intensities (0,05mJ/mm2) and 500 number of shocks (impulses), whereas control cells received no treatment. Cell division and apoptosis are interconnected and regulated by cyclins, kinases, Bax and Opg. We valued the cell viability quantifying by PCR the expressions of these molecules which regulate cell-cycle.

Results: We found an immediate negative effect on cell viability, that occurs with an increase of Bax protein expression after 3 hours of treatment. After a longer time lapse a stimulatory effect on cell proliferation, as reflected by the increase of a G(1)-S phase marker, was observed. In fact, in the following 24, 48 and 72 hours after ESW treatment, we found a stronger association of Cyclin E2 and Cdk2, forming active cyclin E-Cdk2 kinase, compared to untreated cells at the same times. We further explored the molecular mechanism for the ESW induction of osteogenesis: by Real Time PCR an enhancement of Runx2 mRNA, evident 48 hours after the treatment, was found. A link between physical ESW and Runx2 activation has been already demonstrated. ESW-induced O2- production, followed by tyrosine kinase mediated ERK activation and Runx2 activation, resulted in osteogenic cell growth and maturation. Moreover, we analyzed the cytokines RANK-L and OPG osteoblast expression, involved in regulation of osteoclastogenesis. A decrease in RANK-L /OPG ratio was found, perhaps leading to a reduced osteoclastogenesis.

Conclusions: The Shock waves have a repair action on bone and it can been explained by the regulation on osteoclastogenesis by the apoptoic pathway of Bax and Opg.


Orthopaedic Proceedings
Vol. 90-B, Issue SUPP_I | Pages 182 - 182
1 Mar 2008
Quagliarella L Sasanelli N Moretti B Patella V
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Clinical follow-up of hip and knee arthroplasty is not related to objective functional parameters while this is one of the main goal of evidence based medicine. Therefore a functional test was defined in order to correlate clinical and biomechanical data.

The experimental set-up has been presented [1] as well as the test protocol [2]. Three parameters have been analyzed: reaction time (Tr), flight time (Tf) and maximum force (Fmax). The data refer to 21 subjects with hip joint replacement (HRG) and 22 subjects with knee joint replacement (KRG). Tests, were performed before surgery and after one, three and six months. The results were compared with values obtained from a control group of 402 normal subjects.

One months after surgery the performance is lower respect to normal data, both for HRG and KRG. Three months later, there is a partial recover expecially for HRG. At six months follow-up, also the KRG reach better performance. While the performance starting point is higher in the HRG, the percentage recovery is equal in both the groups. During the follow-up also the non operated leg, both for HRG and KRG, shows a progressive changing in its performances, which can increase or decrease, but always it brings at the same level of ability for both legs.

The data suggest that there are different performance and time recover related to the replaced joint (hip or knee) while the total amount of recover is not joint related and there are no differences for laterality. Experimental data correlate with clinical observation; therefore the proposed protocoll seems to be usefull for objective evaluation of joint replacement follow-up. The re-equilibration of the kinematic abilities between the limbs, which cannot be detected by clinical observation, requires further investigation and could be related to neurological integration and less algia limitation.


The Journal of Bone & Joint Surgery British Volume
Vol. 82-B, Issue 3 | Pages 457 - 460
1 Apr 2000
Zambonin G Camerino C Greco G Patella V Moretti B Grano M

We have studied in vitro the effect of a hydroxyapatite (HA) tricalcium phosphate material coated with hepatocyte growth factor (HA-HGF) on cell growth, collagen synthesis and secretion of metalloproteinases (MMPs) by human osteoblasts.

Cell proliferation was stimulated when osteoblasts were incubated with untreated HA and was further increased after exposure to HA-HGF. The uptake of [3H]-proline was increased after treatment with HA. When osteoblasts were exposed to HA-HGF, collagen synthesis was increased with respect to HA. The secretion of MMPs in control cells was undetectable, but in HA and HA-HGF cells MMP 2 and MMP 9 were clearly synthesised.

Our results suggest that HA can promote osteoblast activity and that HGF can further increase its bioactivity.