In vitro and in vivo evaluation of orthopedic interface repair using a tissue scaffold with a continuous hard tissue-soft tissue transition.

Tendon tears produce pain and decrease joint stability; each year, over 1.1 million rotator cuff tendon surgical procedures are performed worldwide. However, surgical success is highly variable, and the inability of the procedure to drive the regeneration of the normal tendon-bone interface has been identified as a key factor in surgical failure. This study focuses on the development, in vitro evaluation, and in vivo assessment of a tissue scaffold derived from bovine cancellous bone with the potential to direct regeneration of a bone-soft tissue interface. The scaffold is a highly porous scaffold with a continuous hard tissue-soft tissue transition that facilitates load transfer across the interface and contains all of the extracellular matrix components of the orthopedic interface. This study demonstrated the in vitro characterization of the mechanical properties and successful in vivo assessment using an ovine model.

Comparison of insertion characteristics of two types of hydroxyapatite-coated and uncoated positive profile transfixation pins in the third metacarpal bone of horses.

OBJECTIVE: To determine the effect of 2 hydroxyapatite pin coatings on heat generated at the bone-pin interface and torque required for insertion of transfixation pins into cadaveric equine third metacarpal bone. SAMPLE POPULATION: Third metacarpal bone pairs from 27 cadavers of adult horses. PROCEDURES: Peak temperature of the bone at the cis-cortex and the hardware and pin at the trans-cortex was measured during insertion of a plasma-sprayed hydroxyapatite (PSHA)-coated, biomimetic hydroxyapatite (BMHA)-coated, or uncoated large animal transfixation pin. End-insertional torque was measured for each pin. The bone-pin interface was examined grossly and histologically for damage to the bone and coating. RESULTS: The BMHA-coated transfixation pins had similar insertion characteristics to uncoated pins. The PSHA-coated pins had greater mean peak bone temperature at the cis-cortex and greater peak temperature at the trans-cortex (70.9 +/- 6.4(o)C) than the uncoated pins (38.7 +/- 8.4(o)C). The PSHA-coated pins required more insertional torque (10,380 +/- 5,387.8 Nmm) than the BMHA-coated pins (5,123.3 +/- 2,296.9 Nmm). Four of the PSHA-coated pins became immovable after full insertion, and 1 gross fracture occurred during insertion of this type of pin. CONCLUSIONS AND CLINICAL RELEVANCE: The PSHA coating was not feasible for use without modification of presently available pin hardware. The BMHA-coated pins performed similarly to uncoated pins. Further testing is required in an in vivo model to determine the extent of osteointegration associated with the BMHA-coated pins in equine bone.

Gentamicin concentrations in synovial fluid obtained from the tarsocrural joints of horses after implantation of gentamicin-impregnated collagen sponges.

OBJECTIVE: To determine synovial fluid gentamicin concentrations and evaluate adverse effects on the synovial membrane and articular cartilage of tarsocrural joints after implantation of a gentamicin-impregnated collagen sponge. ANIMALS: 6 healthy adult mares. PROCEDURES: A purified bovine type I collagen sponge impregnated with 130 mg of gentamicin was implanted in the plantarolateral pouch of 1 tarsocrural joint of each horse, with the contralateral joint used as a sham-operated control joint. Gentamicin concentrations in synovial fluid and serum were determined for 120 hours after implantation by use of a fluorescence polarization immunoassay. Synovial membrane and cartilage specimens were collected 120 hours after implantation and evaluated histologically. RESULTS: Median peak synovial fluid gentamicin concentration of 168.9 microg/mL (range, 115.6 to 332 microg/mL) was achieved 3 hours after implantation. Synovial fluid gentamicin concentrations were < 4 microg/mL by 48 hours. Major histologic differences were not observed in the synovial membrane between control joints and joints implanted with gentamicin-impregnated sponges. Safranin-O fast green stain was not reduced in cartilage specimens obtained from treated joints, compared with those from control joints. CONCLUSIONS AND CLINICAL RELEVANCE: Implantation of a gentamicin-impregnated collagen sponge in the tarsocrural joint of horses resulted in rapid release of gentamicin, with peak concentrations > 20 times the minimum inhibitory concentration reported for common pathogens that infect horses. A rapid decrease in synovial fluid gentamicin concentrations was detected. The purified bovine type I collagen sponges did not elicit substantial inflammation in the synovial membrane or cause mechanical trauma to the articular cartilage.

Safety of, and biological and functional response to, a novel metallic implant for the management of focal full-thickness cartilage defects: Preliminary assessment in an animal model out to 1 year.

Focal full-thickness cartilage lesions of the human medial femoral condyle (MFC) can cause pain and functional impairment. Affected middle-aged patients respond unpredictably to existing treatments and knee arthroplasty may be required, prompting risk of revision. This study assesses the safety of, and biological and functional response to, a metallic resurfacing implant which may delay or obviate the need for traditional arthroplasty. The anatomic contour of the surgically exposed MFC of six adult goats was digitally mapped and an 11 mm diameter full-thickness osteochondral defect was created. An anchor-based Co-Cr resurfacing implant, matching the mapped articular contour, was implanted. Each goat's contralateral unoperated femorotibial joint was used as a control. Postoperative outcome was assessed by lameness examination, radiography, arthroscopy, synoviocentesis, necropsy, and histology up to 26 (n = 3) or 52 (n = 3) weeks. By postoperative week (POW) 4, goats demonstrated normal range of motion, no joint effusion, and only mild lameness in the operated limb. By POW 26 the animals were sound with only occasional very mild lameness. Arthroscopy at POW 14 revealed moderate synovial inflammation and a chondral membrane extending centrally across the implant surface. Radiographs at POWs 14 to 52 implied implant stability in the operated joints, as well as subchondral bone remodeling and mild exostosis formation in the operated and contralateral unoperated joints of some goats. By POW 26, histology revealed new trabecular bone abutting the implant. At POWs 26 and 52 MFC cartilage was metachromatic and intact in the operated and unoperated femorotibial joints. Proximal tibiae of some operated and unoperated limbs demonstrated limited subchondral bone remodeling and foci of articular cartilage fibrillation and thinning. The chondral membrane crossing the prosthesis possessed a metachromatic matrix containing singular and clustered chondrocytes. Our data imply the safety, biocompatibility, and functionality of the implant. Focal articular damage was documented in the operated joints at POWs 26 and 52, but lesions were much reduced over those previously reported in untreated defects. Expanded animal or preclinical human studies are justified.

Nonlinear optical microscopy of articular cartilage.

OBJECTIVE: To assess the ability of nonlinear optical microscopy (NLOM) to image ex vivo healthy and degenerative bovine articular cartilage. METHOD: Fresh bovine femoral-tibial joints were obtained from an abattoir. Articular cartilage specimens were harvested from the tibial plateau. Normal and degenerative specimens were imaged by NLOM and subsequently fixed and processed for histological examination. RESULTS: NLOM provided high resolution images of articular cartilage at varying depths with high sensitivity to tissue morphology and high specificity to tissue components without fixing, sectioning or staining. Spectroscopic segmentation of nonlinear optical signals isolated the collagen matrix from the chondron (chondrocyte and non-collagen pericellular matrix). Images from the superficial zone were consistent with the presence of a matrix composed of both elastin-like and collagen fibers distributed in a depth-dependent morphological arrangement, whereas only collagen was demonstrated in the middle and deep zones. Alterations of collagen matrix associated with advanced degenerative joint disease (fibrocartilage) were observed with NLOM. Individual chondrocytes were imaged and demonstrated intracellular fluorescence consistent with the presence of products of intracellular biochemical processes. CONCLUSION: Thin images of living articular cartilage using NLOM may be obtained with (sub-)cellular resolution at varying depths without fixing, sectioning or staining. Extracellular matrical collagen and chondron may be imaged separately in native tissue using spectrally distinct, endogenous, nonlinear optical signals. NLOM was sensitive to macromolecular composition and pathologic changes in articular cartilage matrix. Advances in instrumentation may lead to the application of NLOM to study articular cartilage in vivo.

Cambium cell stimulation from surgical release of the periosteum.

An autograft of periosteal tissue containing cambium cells has potential to become chondrogenic or osteogenic depending on the regeneration repair strategies. The potential number of harvestable cambium cells diminishes with age. Other factors may be associated with a reduction in the number or variable yields of cambium cells including harvest technique, harvest site location, and the time interval from harvest to implantation. Attempts to increase the number of cambium cells have included improvements in harvesting and handling technique, and expansion of the cells in tissue culture. An "in situ" stimulation and proliferation technique would offer the potential for increasing the number of cambium cells in a cost-effective manner for transplantation without the need for expansion in tissue culture. The hypothesis tested was that surgical release of the periosteum and its deep inner underlying cambium layer by sharply incising through the superficial periosteal fibrous layer down to and scoring the cortical bone surface would increase the number of cambium cells that could be harvested at a later time period. Two techniques for periosteal release were used to stimulate a proliferation of the underlying cambium layer and increase the cambium cells for harvest in skeletally mature goats: (1) sharply scoring all four-sides of the tissue test site perimeter, and (2) sharply scoring only two sides of the tissue test site. The two-sided and four-sided release scoring of the periosteum induced stimulatory responses in the number of cambium cells. In addition, a marked increase in mRNA expression for BMP-2 (p<0.001) was observed within 24 h and remained elevated over baseline values for up to 96 h after this stimulation to the cambium layer.

Effects of continuous intra-articular infusion of gentamicin on synovial membrane and articular cartilage in the tarsocrural joint of horses.

OBJECTIVE: To determine the effects of a continuous intra-articular infusion of gentamicin on the synovial membrane and articular cartilage in the tarsocrural joint of horses. ANIMALS: 6 healthy adult horses. PROCEDURE: A balloon infusion system attached to a catheter placed in the plantarolateral pouch of both tarsocrural joints in each horse was used for continuous gentamicin solution (GM) or balanced electrolyte solution (BES) delivery for 5 days. Cartilage and synovial membrane specimens were collected on day 5 from 3 horses and on day 14 from the remaining 3 horses. Both infused joints from each horse were assessed, using gross evaluation and histologic scoring systems. RESULTS: Significant differences in the histologic scores of synovial membrane specimens between the GM- and BES-treated joints at either 5 or 14 days were not observed. Safranin-O-fast green staining scores were similar between cartilage specimens from GM- and BES-treated joints. Although the synovial membrane histologic scores and safranin-O-fast green staining scores improved from day 5 to 14, the changes in scores were not significant. Loss of synovial intimal cells from villi was found more commonly in sections of synovial membrane from GM-treated joints, compared with BES-treated joints. CONCLUSIONS AND CLINICAL RELEVANCE: Continuous infusion of GM into the tarsocrural joint of horses does not have significant effects on histologic scores of articular cartilage or synovial membrane, compared with those infused with BES. Continuous infusion of GM into the tarsocrural joint of horses for 5 days is an acceptable method for the treatment of septic arthritis.