Zygomatic arch reconstruction with a patient-specific polycaprolactone/beta-tricalcium phosphate scaffold after parosteal osteosarcoma resection in a dog.

OBJECTIVE: To describe the surgical application of a 3D-printing-based, patient-specific, biocompatible polycaprolactone/beta-tricalcium phosphate (PCL/ß-TCP) scaffold to reconstruct the zygomatic arch after tumor resection in a dog. ANIMAL: A 13 year old female spayed Maltese. STUDY DESIGN: Case report METHODS: The dog's presenting complaint was swelling ventral to her right eye. A round mass arising from the caudal aspect of the right zygomatic arch was identified by computed tomography (CT). The histopathologic diagnosis was a low-grade spindle-cell tumor. Surgical resection was planned to achieve 5 mm margins. A patient-specific osteotomy guide and polycaprolactone/beta-tricalcium phosphate (PCL/ß-TCP) scaffold were produced. Osteotomy, including 30% of total zygomatic arch length, was performed using an oscillating saw aligned with the guide. The scaffold was placed in the defect. Parosteal osteosarcoma was diagnosed based on histopathological examination. Excision was complete, with the closest margin measuring 0.3 mm. RESULTS: Mild epiphora, due to surgical site swelling, subsided after 20 days. Tissue formation within and around the porous scaffold was noted on CT 10 months postoperatively, with no evidence of metastasis or local recurrence. Facial conformation appeared symmetrical, and no complications were noted 16 months postoperatively. CONCLUSION: The use of a 3D-printing-based, patient-specific, biocompatible PCL/ß-TCP scaffold successfully restored the structure and function of the zygomatic arch without complications, even following wide zygomectomy for complete tumor removal.

Influence of a novel scaffold composed of polyurethane, hydroxyapatite, and decellularized bone particles on the healing of fourth metacarpal defects in mares.

OBJECTIVE: To determine the effect of a novel scaffold, designed for use in bone regeneration, on healing of splint bone segmental defects in mares. STUDY DESIGN: In vivo experimental study. SAMPLE POPULATION: Five adult mares (4-10 years old; mean weight, 437.7 kg ± 29 kg). METHODS: Bilateral 2-cm full-thickness defects were created in the fourth metacarpal bones (MCIV) of each horse. Each defect was randomly assigned to either a novel scaffold treatment (n = 5) or an untreated control (n = 5). The scaffold was composed of polyurethane, hydroxyapatite, and decellularized bone particles. Bone healing was assessed for a period of 60 days by thermography, ultrasonography, radiography, and computed tomography (CT). Biopsies of each defect were performed 60 days after surgery for histological evaluation. RESULTS: On the basis of radiographic analysis, scaffold-treated defects had greater filling (67.42% ± 26.7%) compared with untreated defects (35.88% ± 32.7%; P = .006). After 60 days, CT revealed that the density of the defects treated with the scaffolds (807.80 ± 129.6 Hounsfield units [HU]) was greater than density of the untreated defects (464.80 ± 81.3 HU; P = .004). Evaluation of histology slides provided evidence of bone formation within an average of 9.43% ± 3.7% of the cross-sectional area of scaffolds in contrast to unfilled defects in which connective tissue was predominant throughout the biopsy specimens. CONCLUSION: The novel scaffold was biocompatible and supported bone formation within the MCIV segmental defects. CLINICAL SIGNIFICANCE: This novel scaffold offers an effective option for filling bone voids in horses when support of bone healing is indicated.

Corneoconjunctival transposition with and without ACell® for deep corneal ulcer repair in 18 dogs.

OBJECTIVE: To document a case series using corneoconjunctival transposition (CCT) surgery with and without bioscaffolding matrix (ACell® ) to repair deep corneal ulcers and perforations in dogs. ANIMALS STUDIED: Eighteen dogs of various breeds that presented with deep or perforating corneal ulcers. PROCEDURES: Corneoconjunctival transposition grafts with or without ACell® were sutured using a simple interrupted 8-0 or 9-0 polyglactin 910 pattern. RESULTS: A total of eighteen dogs (19 eyes) were diagnosed with deep corneal ulcers (n = 7) and perforating corneal ulcers (n = 12). A CCT was performed in all eyes, with ten of them additionally receiving an ACell® graft. The majority of lesions were located axially in 14/19 (81%) eyes. Grafts were harvested from dorsal (n = 8), temporal (n = 5), ventral (n = 4), or nasal (n = 2) quadrants. Brachycephalic breeds (13/18) were over-represented. Keratoconjunctivitis sicca was present in 10/19 eyes (52.6%). Bacterial isolates were cultured from 8/19 eyes. Post-operative therapy included topical antibiotics, plasma, cycloplegics, oral antibiotics, and oral nonsteroidal anti-inflammatory drugs. CCT integration with and without ACell® occurred at a median of 20 days (range 7-38 days) post-operatively with no significant difference between groups. Median follow-up time was 188 days. Short-term post-operative complications included granulation tissue formation (19/19), corneal edema (4/19), graft retraction (4/19), and anterior synechia (1/19). Long-term complications in 14 eyes with follow-up >30 days included superficial corneal pigmentation (6/14) and epithelial inclusion cysts (5/14). Two eyes were nonvisual at last follow-up due to cataract formation. CONCLUSIONS: Corneoconjunctival transposition with ACell® can be utilized for corneal ulcer repair in dogs.

In vivo application of decellularized rat colon and evaluation of the engineered scaffolds following 9 months of follow-up.

The aim of this study was to investigate the rat small intestine mesentery and colon as natural bio-reactors for rat colon-derived scaffolds. We decellularized eight whole rat colons by a perfusion-based protocol using 0.1% sodium dodecyl sulfate for 24 hr. The provided bio-scaffolds were examined by histological staining, scanning electron microscopy, and collagen and sulfated glycosaminoglycan quantification. Subsequently, we implanted 4 cm segments of the provided bio-scaffolds into two groups of animal models comprising tissue grafting into the mesenteric tissue (n: 10) and end-to-end anastomosis (n: 10) to the colon of host rats. Following 9 months of follow-up, we harvested the grafts and performed histological and immunohistochemical studies as well as real-time PCR evaluation for telomerase activity of the samples. Histological staining, scanning electron microscopy and protein content evaluation of the acellular tissues confirmed the complete removal of the cellular components and preservation of the extracellular matrix. Histopathological assessment of the implanted scaffolds was suggestive of a regenerative process in both groups. Moreover, immunohistochemical analysis of the samples confirmed the presence of smooth muscle cells, endothelial progenitor cells, and neural elements in both groups of grafted scaffolds. Our data confirmed the recellularization of the acellular colon grafts in both groups after 9 months of follow up. Also, the implanted tissues demonstrated different characteristics based on their implantation location. The outcomes of this investigation illustrate the capability of acellular tissues for in vivo application and regeneration.

Development and assessment of a novel ex vivo corneal culture technique involving an agarose-based dome scaffold for use as a model of in vivo corneal wound healing in dogs and rabbits.

OBJECTIVE: To develop and assess a novel ex vivo corneal culture technique involving an agarose-based dome scaffold (ABDS) for use as a model of in vivo corneal wound healing in dogs and rabbits. SAMPLE: Corneas from clinically normal dogs (paired corneas from 8 dogs and 8 single corneas) and rabbits (21 single corneas). PROCEDURES: 8 single dog corneas (DCs), 1 DC from each pair, and 10 rabbit corneas (RCs) were wounded with an excimer laser; 1 DC from each pair and 11 RCs remained unwounded. Corneas were cultured for 21 days on ABDSs (8 pairs of DCs and all RCs) or on flat-topped scaffolds (8 single DCs). The surface area of corneal fluorescein retention was measured every 6 (DCs) or 12 (RCs) hours until full corneal epithelialization was detected. Changes in corneal clarity were evaluated at 0, 7, 14, and 21 days. RESULTS: Median time to full epithelialization for wounded dog and rabbit corneas was 48 and 60 hours, respectively; among wounded DCs, time to full epithelization did not differ by scaffold type. After 21 days of culture on ABDSs, all DCs and RCs that epithelialized developed a circular, diffuse, cloud-like pattern of optical haze, whereas DCs cultured on flat-topped scaffolds developed a focal, crater-like region of optical haze. All corneas on the ABDSs maintained convex curvature throughout the study. CONCLUSIONS AND CLINICAL RELEVANCE: Wounded ex vivo DCs and RCs cultured on ABDSs reliably epithelialized, formed optical haze (consistent with in vivo wound healing), and maintained convex curvature. This culture technique may be adaptable to other species.

New clinical application of three-dimensional-printed polycaprolactone/ß-tricalcium phosphate scaffold as an alternative to allograft bone for limb-sparing surgery in a dog with distal radial osteosarcoma.

Limb-sparing surgery is one of the surgical options for dogs with distal radial osteosarcoma (OSA). This case report highlights the novel application of a three-dimensional (3D)-printed patient-specific polycaprolactone/ß-tricalcium phosphate (PCL/ß-TCP) scaffold in limb-sparing surgery in a dog with distal radial OSA. The outcomes evaluated included postoperative gait analysis, complications, local recurrence of tumor, metastasis, and survival time. Post-operative gait evaluation showed significant improvement in limb function, including increased weight distribution and decreased asymmetry. The implant remained well in place and increased bone opacity was observed between the host bone and the scaffold. There was no complication due to scaffold or surgery. Significant improvement in limb function and quality of life was noted postoperatively. Local recurrence and pulmonary metastasis were identified at 8 weeks postoperatively. The survival time from diagnosis of OSA to death was 190 days. The PCL/ß-TCP scaffold may be an effective alternative to cortical allograft in limb-sparing surgery for bone tumors.

Zinc silicate mineral-coated scaffold improved in vitro osteogenic differentiation of equine adipose-derived mesenchymal stem cells.

In current study we aimed to coat the PLLA scaffold with zinc (Zn) silicate mineral nanoparticles. Then, using equine adipose-derived stem cells (ASCs) we intended to compare the osteogenic induction potency of Zn silicate mineral-coated PLLA scaffold with uncoated PLLA scaffold and tissue culture plastic (TCPS). Adipose tissues were collected from 3 horses, and isolation of ASCs was achieved by enzymatic digestion. PLLA scaffold was successfully prepared using a phase separation method and coated with Zn silicate mineral nanoparticles. The coating efficiency was then characterized by scanning electron microscopy and further evaluated with the application of fourier transform infrared microscopic imaging. Viability and growth characteristics of ASCs on TCPS, uncoated and coated PLAA scaffolds were investigated by MTT assay. Alizarin Red staining was performed for determination of calcium deposition following the osteogenic induction. Furthermore, other common osteogenic markers such as alkaline phosphatase (ALP) activity, calcium content, as well as osteogenic (Runx2, ALP, osteonectin, and collagen I) marker genes were also evaluated. Our data showed that Zn silicate mineral nanoparticles was coated successfully on PLLA scaffold and such scaffold had no detrimental effect on cell growth rate as indicated by MTT assay. Moreover, ASCs that differentiated on Zn silicate mineral-coated PLLA scaffold indicated higher ALP activity, more calcium content, and higher expression of bone-related genes than that on uncoated PLLA scaffold and TCPS. Adequate proliferation rate and higher expression of osteogenic markers of stem cells, provides this scaffold as a suitable substrate to support proliferation and differentiation of ASCs in equine.

Application of combined porous tantalum scaffolds loaded with bone morphogenetic protein 7 to repair of osteochondral defect in rabbits.

PURPOSE: Porous tantalum (PT) has been widely used in orthopaedic applications for low modulus of elasticity, excellent biocompatibility, and the microstructures similar to cancellous bone. In order to improve the biological activity of PT, biologically active factors can be combined with the material. The purpose of this study was to investigate if bone morphogenetic protein 7 (BMP-7) modifications could enhance the repairing of cartilage of PT in osteochondral defect in medial femoral condyle of rabbits. METHODS: A cylindrical osteochondral defect model was created on the animal medial femoral condyle of and filled as follows: PT modified with BMP-7 for MPT group, non-modified PT for the PT group, while no implants were used for the blank group. The regenerated osteochondral tissue was assessed and analyzed by histological observations at four, eight and 16 weeks post-operation and evaluated in an independent and blinded manner by five different observers using a histological score. Osteochondral and subchondral bone defect repair was assessed by micro-CT scan at 16 weeks post-operation, while the biomechanical test was performed at 16 weeks post-operation. RESULTS: Briefly, higher overall histological score was observed in the MPT group compared to PT group. Furthermore, more new osteochondral tissue and bone formed at the interface and inside the inner pores of scaffolds of the MPT group compared to PT group. Additionally, the micro-CT data suggested that the new bone volume fractions and the quantity and quality of trabecular bone, as well as the maximum release force of the bone, were higher in the MPT group compared to PT group. CONCLUSIONS: We demonstrated that the applied modified PT with BMP-7 promotes excellent subchondral bone regeneration and may serve as a novel approach for osteochondral defects repair.

Three-Dimensional Printing-based Reconstruction of a Maxillary Bone Defect in a Dog Following Tumor Removal.

Three-dimensional (3D) printing has been applied extensively not only in human, but also veterinary medicine. However, the technique is still used in the clinical area for a surgical plan or education prior to surgery. Thus, we report a case of reconstruction after tumor removal surgery with the use of a 3D-printed scaffold. A 12-year-old female mixed dog had a left caudal maxillary mass. Based on computed tomography images, a defect was confirmed on the maxillary bone due to the oral mass, and a surgical plan was designed to remove the oral mass and graft the 3D printed scaffold. Customized polycaprolactone/ beta-tracalciumphosphate (PCL/ß-TCP) scaffold was fabricated using the micro-extrusion-based 3D printer. In the operation, after the removal of the oral mass, the scaffold was grafted onto the defect site. At follow-up, 8 months after surgery, the result was successful without any special problems in the periodic CT scans and oral examinations. This case is believed to be the first case of reconstruction by using a 3D printed scaffold in the maxillary bone defect, and this 3D printing technique is thought to be very helpful for veterinary patients with bone defects and several other diseases.

Modified penetrating keratoplasty with Acell® bioscaffold implant in seven horses with deep full-thickness corneal stromal abscess.

OBJECTIVE: To describe and evaluate a modified penetrating keratoplasty technique utilizing ACell® for management of equine deep stromal or full-thickness corneal stromal abscesses (SA). METHODS: Cases presenting to the University of Georgia Ophthalmology service for surgical management of SA necessitating penetrating keratoplasty (PK) were included in the study population. Surgery entailed the use of an ACell® disk sutured within the deep level of a stepped full-thickness corneal incision with an overlying conjunctival pedicle flap placed in the superficial step incision. Patients were evaluated for success as defined by a comfortable, visual outcome. RESULTS: Surgery was performed in seven horses. Conjunctival flap incorporation and globe retention occurred in all patients. Functional vision was maintained in six of seven eyes (85.7%) at last follow-up examination (mean of 87.6 days [range 41-251 days]). Mean size of ACell® implant was six millimeters (range 4-8 mm). Postoperative complications included moderate to severe anterior uveitis (n = 2), diffuse keratitis (n = 1), incipient cataract formation (n = 3), and anterior and posterior synechiae (n = 1). CONCLUSIONS: This technique is a viable option for treatment of equine SA requiring PK. The use of bioscaffold implant is an alternative to frozen and fresh donor cornea transplantation.

Comparison of four methods for generating decellularized equine synovial extracellular matrix.

OBJECTIVE To evaluate 4 methods for generating decellularized equine synovial extracellular matrix. SAMPLE Villous synovium harvested from the femoropatellar and medial femorotibial joints of 4 healthy adult horses < 7 years of age. Synovial samples were frozen (-80°C) until used. PROCEDURES Synovial samples were thawed and left untreated (control) or decellularized with 1 of 4 methods (15 samples/horse/method): incubation in 0.1% peracetic acid (PAA), incubation in 0.1% PAA twice, incubation in 1% Triton X-100 followed by incubation in DNase, and incubation in 2M NaCl followed by incubation in DNase. Control and decellularized samples were examined for residual cells, villous integrity, and collagen structure and integrity by means of histologic examination and scanning electron microscopy; cell viability was evaluated by means of culture and exclusion staining. Decellularization efficiency was assessed by testing for DNA content and DNA fragment size. RESULTS Incubation in PAA once preserved the synovial villous architecture, but resulted in high DNA content and retention of large (> 25,000 base pair) DNA fragments. Incubation in Triton and incubation in NaCl resulted in low DNA content and short (< 200 base pair) DNA fragments, but destroyed the synovial villous architecture. Incubation in PAA twice resulted in low DNA content and short DNA fragments while retaining the synovial villous architecture. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that of the methods evaluated, incubation in 0.1% PAA twice was the best method for generating decellularized equine synovial extracellular matrix.

Isolation, cultivation and characterisation of pigeon osteoblasts seeded on xenogeneic demineralised cancellous bone scaffold for bone grafting.

Avian osteoblasts have been isolated particularly from chicken embryo, but data about other functional tissue sources of adult avian osteoblast precursors are missing. The method of preparation of pigeon osteoblasts is described in this study. We demonstrate that pigeon cancellous bone derived osteoblasts have particular proliferative capacity in vitro in comparison to mammalian species and developed endogenous ALP. Calcium deposits formation in vitro was confirmed by alizarin red staining. Only a few studies have attempted to investigate bone grafting and treatment of bone loss in birds. Lack of autologous bone grafts in birds has prompted investigation into the use of avian xenografts for bone augmentation. Here we present a method of xenografting of ostrich demineralised cancellous bone scaffold seeded with allogeneic adult pigeon osteoblasts. Ostrich demineralised cancellous bone scaffold supported proliferation of pigeon osteoblasts during two weeks of co - cultivation in vitro. Scanning electron microscopy demonstrated homogeneous adult pigeon osteoblasts attachment and distribution on the surface of xenogeneic ostrich demineralised cancellous bone. Our preliminary in vitro results indicate that demineralised cancellous bone from ostrich tibia could provide an effective biological support for growth and proliferation of allogeneic osteoblasts derived from cancellous bone of pigeons.

Comparison of meniscal fibrochondrocyte and synoviocyte bioscaffolds toward meniscal tissue engineering in the dog.

Tissue engineering is a promising field of study toward curing the meniscal deficient stifle; however the ideal cell type for this task is not known. We describe here the extraction of synoviocytes and meniscal fibrochondrocytes from arthroscopic debris from six dogs, which were cultured as tensioned bioscaffolds to synthesize meniscal-like fibrocartilage sheets. Despite the diseased status of the original tissues, synoviocytes and meniscal fibrochondrocytes had high viability at the time of removal from the joint. Glycosaminoglycan and collagen content of bioscaffolds did not differ. Meniscal fibrochondrocyte bioscaffolds contained more type II collagen, but collagen deposition was disorganized, with only 30-40% of cells viable. The collagen of synoviocyte bioscaffolds was organized into sheets and bands and 80-90% of cells were viable. Autologous, diseased meniscal fibrochondrocytes and synoviocytes are plausible cell sources for future meniscal tissue engineering research, however cell viability of meniscal fibrochondrocytes in the tensioned bioscaffolds was low.

Comparison of growth factor treatments on the fibrochondrogenic potential of canine fibroblast-like synoviocytes for meniscal tissue engineering.

OBJECTIVE: To determine the in vitro effects of differing growth factor treatments on the fibrochondrogenic potential of fibroblast-like synoviocytes from cruciate ligament deficient femorotibial joints of dogs. STUDY DESIGN: In vitro study. SAMPLE POPULATION: Synoviocytes from dogs (n = 8) with naturally occurring cruciate ligament insufficiency. METHODS: Synoviocytes were cultured in monolayer and synthesized into tensioned synoviocyte bioscaffolds (TSB) suspended in media containing TGF-ß3, or FGF-2, TGF-ß1, and IGF-I. The 1,9-dimethylmethylene blue (DMMB) assay and toluidine blue stain assessed glycosaminoglycan content; hydroxyproline assay, and collagen I and II immunohistochemistry assessed collagen content. Biomechanical properties were determined by materials testing/force-deformation curves. RESULTS: All tissue cultures formed tensioned fibrous tissue-like constructs. Mean tissue cellularity and cellular viability was significantly greater in the triple growth factor-treated TSB by 0.09% and 44%, respectively. Percentage collagen content, and relative gene expression for collagen I, II, and aggrecan was not significantly different between groups. Median percentage of GAG content was significantly greater in triple growth factor-treated TSB by 1.6%. Biomechanical properties were not different in compression. Triple growth factor-treated TSB were significantly stronger in toughness, peak load to failure, and stiffness in tension. CONCLUSIONS: TGF-ß3 cultured bioscaffolds failed to outperform triple growth factor-treated TSB. Architectural extracellular matrix (ECM) organization and cellularity likely explained the differences between groups. TGF-ß3 alone cannot be recommended at this time for in vitro formation of autologous fibrocartilage bioscaffolds for meniscal deficiency.

Application of a 3D printed customized implant for canine cruciate ligament treatment by tibial tuberosity advancement.

Fabrication of customized implants based on patient bone defect characteristics is required for successful clinical application of bone tissue engineering. Recently a new surgical procedure, tibial tuberosity advancement (TTA), has been used to treat cranial cruciate ligament (CrCL) deficient stifle joints in dogs, which involves an osteotomy and the use of substitutes to restore the bone. However, limitations in the use of non-biodegradable implants have been reported. To overcome these limitations, this study presents the development of a bioceramic customized cage to treat a large domestic dog assigned for TTA treatment. A cage was designed using a suitable topology optimization methodology in order to maximize its permeability whilst maintaining the structural integrity, and was manufactured using low temperature 3D printing and implanted in a dog. The cage material and structure was adequately characterized prior to implantation and the in vivo response was carefully monitored regarding the biological response and patient limb function. The manufacturing process resulted in a cage composed of brushite, monetite and tricalcium phosphate, and a highly permeable porous morphology. An overall porosity of 59.2% was achieved by the combination of a microporosity of approximately 40% and a designed interconnected macropore network with pore sizes of 845 µm. The mechanical properties were in the range of the trabecular bone although limitations in the cage's reliability and capacity to absorb energy were identified. The dog's limb function was completely restored without patient lameness or any adverse complications and also the local biocompatibility and osteoconductivity were improved. Based on these observations it was possible to conclude that the successful design, fabrication and application of a customized cage for a dog CrCL treatment using a modified TTA technique is a promising method for the future fabrication of patient-specific bone implants, although clinical trials are required.

Growth factor treated tensioned synoviocyte neotissues: towards meniscal bioscaffold tissue engineering.

Meniscal injury is a common cause of osteoarthritis, pain, and disability in dogs and humans, but tissue-engineered bioscaffolds could be a treatment option for meniscal deficiency. The objective of this study was to compare meniscus-like matrix histology, composition, and biomechanical properties of autologous tensioned synoviocyte neotissues (TSN) treated with fetal bovine serum (TSNfbs) or three chondrogenic growth factors (TSNgf). Fourth passage canine synoviocytes from 10 dogs were grown in hyperconfluent monolayer culture, formed into TSN, and then cultured for 3 weeks with 17.7% FBS or three human recombinant TSNgf (bFGF, TGF-ß1, and IGF-1). Cell viability was determined with laser microscopy. Histological architecture and the composition of fibrocartilage matrix were evaluated in TSN by staining tissues for glycosaminoglycan (GAG), α-smooth muscle actin, and collagen 1 and 2; quantifying the content of GAG, DNA, and hydroxyproline; and measuring the gene expression of collagens type 1α and 2α, the GAG aggrecan, and transcription factor Sry-type Homeobox Protein-9 (SOX9). Biomechanical properties were determined by materials testing force-deformation curves. The TSN contained components and histological features of mensical fibrocartilage extracellular matrix. Growth factor-treated TSN had higher DNA content but lower cell viability than TSNfbs. TSNgf had greater fibrocartilage-like matrix content (collagen 2 and GAG content with increased collagen 2α and SOX9 gene expression). Additionally, TSNgf collagen was more organized histologically and so had greater tensile biomechanical properties. The results indicate the potential of TSN when cultured with growth factors as implantable bioscaffolds for the treatment of canine meniscal deficiency.

In vitro synthesis of tensioned synoviocyte bioscaffolds for meniscal fibrocartilage tissue engineering.

BACKGROUND: Meniscal injury is a common cause of lameness in the dog. Tissue engineered bioscaffolds may be a treatment option for meniscal incompetency, and ideally would possess meniscus- like extracellular matrix (ECM) and withstand meniscal tensile hoop strains. Synovium may be a useful cell source for meniscal tissue engineering because of its natural role in meniscal deficiency and its in vitro chondrogenic potential. The objective of this study is to compare meniscal -like extracellular matrix content of hyperconfluent synoviocyte cell sheets ("HCS") and hyperconfluent synoviocyte sheets which have been tensioned over wire hoops (tensioned synoviocyte bioscaffolds, "TSB") and cultured for 1 month. RESULTS: Long term culture with tension resulted in higher GAG concentration, higher chondrogenic index, higher collagen concentration, and type II collagen immunoreactivity in TSB versus HCS. Both HCS and TSB were immunoreactive for type I collagen, however, HCS had mild, patchy intracellular immunoreactivity while TSB had diffuse moderate immunoreactivity over the entire bisocaffold. The tissue architecture was markedly different between TSB and HCS, with TSB containing collagen organized in bands and sheets. Both HCS and TSB expressed alpha smooth muscle actin and displayed active contractile behavior. Double stranded DNA content was not different between TSB and HCS, while cell viability decreased in TSB. CONCLUSIONS: Long term culture of synoviocytes with tension improved meniscal- like extra cellular matrix components, specifically, the total collagen content, including type I and II collagen, and increased GAG content relative to HCS. Future research is warranted to investigate the potential of TSB for meniscal tissue engineering.

Equine articular chondrocytes on MACT scaffolds for cartilage defect treatment.

Treatment of cartilage defects poses challenging problems in human and veterinary medicine, especially in horses. This study examines the suitability of applying scaffold materials similar to those used for human cartilage regeneration on equine chondrocytes. Chondrocytes gained from biopsies of the talocrural joint of three horses were propagated in 2D culture and grown on two different scaffold materials, hyaluronan (HYAFF®) and collagen (BioGide®), and evaluated by light and electron microscopy. The equine chondrocytes developed well in both types of materials. They were vital and physiologically highly active. On the surface of the scaffolds, they formed cell multilayers. Inside the hyaluronan web, the chondrocytes were regularly distributed and spanned the large scaffold fibre distances by producing their own matrix sheath. Half-circle-like depressions occasionally found in the cell membrane were probably related to movement on the flexible matrix sheath. Inside the dense collagen scaffold, only single cells were found. They passed through the scaffold strands by cell shape adaptation. This study showed that the examined scaffold materials can be used for equine chondrocyte cultivation. Chondrocytes tend to form multilayers on the surface of both, very dense and very porous scaffolds, and have strategies to span between and move in large gaps.