RESUMO
One of the most challenging aspects of developing advanced cell therapy products (CTPs) is defining the mechanism of action (MOA), potency and efficacy of the product. This perspective examines these concepts and presents helpful ways to think about them through the lens of metrology. A logical framework for thinking about MOA, potency and efficacy is presented that is consistent with the existing regulatory guidelines, but also accommodates what has been learned from the 27 US FDA-approved CTPs. Available information regarding MOA, potency and efficacy for the 27 FDA-approved CTPs is reviewed to provide background and perspective. Potency process and efficacy process charts are introduced to clarify and illustrate the relationships between six key concepts: MOA, potency, potency test, efficacy, efficacy endpoint and efficacy endpoint test. Careful consideration of the meaning of these terms makes it easier to discuss the challenges of correlating potency test results with clinical outcomes and to understand how the relationships between the concepts can be misunderstood during development and clinical trials. Examples of how a product can be "potent but not efficacious" or "not potent but efficacious" are presented. Two example applications of the framework compare how MOA is assessed in cell cultures, animal models and human clinical trials and reveals the challenge of establishing MOA in humans. Lastly, important considerations for the development of potency tests for a CTP are discussed. These perspectives can help product developers set appropriate expectations for understanding a product's MOA and potency, avoid unrealistic assumptions and improve communication among team members during the development of CTPs.
Assuntos
Terapia Baseada em Transplante de Células e Tecidos , Humanos , Terapia Baseada em Transplante de Células e Tecidos/métodos , Animais , Resultado do Tratamento , United States Food and Drug Administration , Estados Unidos , Ensaios Clínicos como AssuntoRESUMO
Large mandibular defects are clinically challenging to reconstruct due to the complex anatomy of the jaw and the limited availability of appropriate tissue for repair. We envision leveraging current advances in fabrication and biomaterials to create implantable devices that generate bone within the patients themselves suitable for their own specific anatomical pathology. The in vivo bioreactor strategy facilitates the generation of large autologous vascularized bony tissue of customized geometry without the addition of exogenous growth factors or cells. To translate this technology, we investigated its success in reconstructing a mandibular defect of physiologically relevant size in sheep. We fabricated and implanted 3D-printed in vivo bioreactors against rib periosteum and utilized biomaterial-based space maintenance to preserve the native anatomical mandibular structure in the defect site before reconstruction. Nine weeks after bioreactor implantation, the ovine mandibles were repaired with the autologous bony tissue generated from the in vivo bioreactors. We evaluated tissues generated in bioreactors by radiographic, histological, mechanical, and biomolecular assays and repaired mandibles by radiographic and histological assays. Biomaterial-aided mandibular reconstruction was successful in a large superior marginal defect in five of six (83%) sheep. Given that these studies utilized clinically available biomaterials, such as bone cement and ceramic particles, this strategy is designed for rapid human translation to improve outcomes in patients with large mandibular defects.
Assuntos
Substitutos Ósseos , Mandíbula , Traumatismos Mandibulares , Periósteo , Impressão Tridimensional , Engenharia Tecidual , Animais , Reatores Biológicos , Feminino , Mandíbula/metabolismo , Mandíbula/patologia , Traumatismos Mandibulares/metabolismo , Traumatismos Mandibulares/patologia , Traumatismos Mandibulares/terapia , Periósteo/metabolismo , Periósteo/patologia , OvinosRESUMO
Sessions included an overview of past cell therapy (CT) conferences sponsored by the International Alliance for Biological Standardization (IABS). The sessions highlighted challenges in the field of human pluripotent stem cells (hPSCs) and also addressed specific points on manufacturing, bioanalytics and comparability, tumorigenicity testing, storage, and shipping. Panel discussions complemented the presentations. The conference concluded that a range of new standardization groups is emerging that could help the field, but ways must be found to ensure that these efforts are coordinated. In addition, there are opportunities for regulatory convergence starting with a gap analysis of existing guidelines to determine what might be missing and what issues might be creating divergence. More specific global regulatory guidance, preferably from WHO, would be welcome. IABS and the California Institute for Regenerative Medicine (CIRM) will explore with stakeholders the development of a practical and innovative road map to support early CT product (CTP) developers.
Assuntos
Terapia Baseada em Transplante de Células e Tecidos , Células-Tronco Pluripotentes , Testes de Carcinogenicidade , Guias como Assunto , Humanos , Controle de Qualidade , Medicina RegenerativaRESUMO
On May 22, 2017, the National Institutes of Health (NIH)/National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS) hosted a roundtable on "Innovative Treatments for Enthesis Repair." A summary of the roundtable discussion, as well as a list of the extramural participants, can be found at https://www.niams.nih.gov/about/meetings-events/roundtables/roundtable-innovative-treatments-enthesis-repair. This paper reviews the challenges and opportunities for developing effective treatment strategies for enthesis repair that were identified at the roundtable discussion.
Assuntos
Osso e Ossos/fisiologia , Traumatismos dos Tendões/fisiopatologia , Cicatrização/fisiologia , HumanosRESUMO
The article summarizes the collective views expressed at the fourth session of the workshop Tissue Engineering--the Next Generation, which was devoted to the translation of results of tissue engineering research into applications. Ernst Hunziker described the paradigm of a dual translational approach, and argued that tissue engineering should be guided by the dimensions and physiological setting of the bodily compartment to be repaired. Myron Spector discussed collagen-glycosaminoglycan (GAG) scaffolds for musculoskeletal tissue engineering. Jeanette Libera focused on the biological and clinical aspects of cartilage tissue engineering, and described a completely autologous procedure for engineering cartilage using the patient's own chondrocytes and blood serum. Arthur Gertzman reviewed the applications of allograft tissues in orthopedic surgery, and outlined the potential of allograft tissues as models for biological and medical studies. Savio Woo discussed a list of functional tissue engineering approaches designed to restore the biochemical and biomechanical properties of injured ligaments and tendons to be closer to that of the normal tissues. Specific examples of using biological scaffolds that have chemoattractants as well as growth factors with unique contact guidance properties to improve their healing process were shown. Anthony Ratcliffe discussed the translation of the results of research into products that are profitable and meet regulatory requirements. Michael Lysaght challenged the proposition that commercial and clinical failures of early tissue engineering products demonstrate a need for more focus on basic research. Arthur Coury described the evolution of tissue engineering products based on the example of Genzyme, and how various definitions of success and failure can affect perceptions and policies relative to the status and advancement of the field of tissue engineering.
Assuntos
Engenharia Tecidual/métodos , Engenharia Tecidual/tendências , Animais , Cartilagem Articular/fisiologia , Humanos , Engenharia Tecidual/normas , Cicatrização/fisiologiaRESUMO
BACKGROUND: High levels of gamma irradiation are required to eliminate the risk of bacterial and viral transmission during implantation of musculoskeletal allografts. The effects of high levels of gamma irradiation on anterior cruciate ligament allograft biomechanics are still not known. HYPOTHESIS: High-dose gamma irradiation (4 Mrad) adversely affects anterior cruciate ligament allograft biomechanics at surgery and at 6 months after surgery and affects biochemistry at 6 months. STUDY DESIGN: Controlled laboratory study. METHODS: Bilateral anterior cruciate ligament reconstructions were performed in 18 adult goats, with one knee receiving an irradiated patellar tendon allograft (4 Mrad) and the other receiving a frozen control allograft (0 Mrad). In 6 recipients (time zero group), graft pairs were tested immediately after sacrifice, and load relaxation of the femur-allograft-tibia preparation was measured during cyclic anterior displacement. Twelve recipients received bilateral anterior cruciate ligament reconstructions, staged 2 months apart, and were sacrificed a mean of 6 months postoperatively. Load relaxation and tensile failure testing were performed, followed by allograft biochemistry assessment. RESULTS: At time zero, irradiated grafts showed less load relaxation than did contralateral controls, but by 6 months, the trend had reversed because of decreases in control graft relaxation, with no changes in irradiated graft relaxation. By 6 months, irradiated grafts showed lower stiffness and maximum force compared to controls but no differences in modulus, maximum stress, or biochemistry. CONCLUSION: High levels of gamma irradiation affect anterior cruciate ligament allograft subfailure viscoelastic and structural properties but not material or biochemical properties over time. CLINICAL RELEVANCE: Although high levels of gamma irradiation may inactivate infectious agents, this treatment is not a feasible clinical option because of altered allograft biomechanics.
Assuntos
Ligamento Cruzado Anterior/cirurgia , Enxerto Osso-Tendão Patelar-Osso , Raios gama , Ligamento Patelar/efeitos da radiação , Animais , Feminino , Glicosaminoglicanos/análise , Cabras , Hidroxiprolina/análise , Modelos Animais , Ligamento Patelar/química , Ligamento Patelar/fisiologia , Proteoglicanas/análise , Distribuição Aleatória , Esterilização/métodos , Resistência à Tração , Fatores de Tempo , Transplante Homólogo , Suporte de CargaRESUMO
The inferior glenohumeral ligament (IGHL) was investigated by correlating the biomechanical properties, biochemical composition, and histologic morphology of its 3 anatomic regions (superior band, anterior axillary pouch, and posterior axillary pouch) in 8 human cadaveric shoulders. The overall biochemical composition of the IGHL appeared similar to other ligaments, with average water content of 80.9 +/- 2.5%, collagen content of 80.0 +/- 9.2%, and crosslinks of 0.715 +/- 0.13 mol/mol collagen. The proteoglycan content was highest in the superior band (2.73 +/- 0.7 mg/g dry weight) and may, in part, explain its viscoelastic behavior. Histologic analysis demonstrated longitudinally organized fiber bundles that were more uniform in the mid-substance but more interwoven and less uniformly oriented near the insertion sites. The superior band had the most pronounced fiber bundle interweaving, while crimping was more evident in the anterior axillary pouch. Elastin was identified in each of the regions. Tensile testing demonstrated a trend toward higher ultimate tensile stress (16.9 +/- 7.9 MPa) and tensile modulus (130.3 +/- 47.9 MPa) in the superior band compared to the axillary pouch. The mean ultimate tensile strain of the IGHL was 16.8 +/- 4.6%. These complex IGHL properties may help to explain its unique functions in stabilizing the shoulder in different arm positions and at different rates of loading, including the failure patterns seen clinically, as in Bankart lesions (insertion site) versus capsular stretching (ligament substance).
Assuntos
Ligamentos Articulares , Articulação do Ombro , Idoso , Fenômenos Biomecânicos , Cadáver , Humanos , Ligamentos Articulares/química , Ligamentos Articulares/patologia , Ligamentos Articulares/fisiologia , Pessoa de Meia-IdadeRESUMO
There is a clear discrepancy between the growth of cell therapy and tissue engineering research in orthopaedics over the last two decades and the number of approved clinical therapies and products available to patients. At the 2015 annual meeting of the Orthopaedic Research Society, a workshop was held to highlight important considerations from the perspectives of an academic scientist, clinical researcher, and industry representative with the aim of helping researchers to successfully translate their ideas into clinical and commercial reality. Survey data acquired from workshop participants indicated an overall positive opinion on the future potential of cell-based therapies to make a significant contribution to orthopaedic medicine. The survey also indicated an agreement on areas requiring improvement in the development of new therapies, specifically; increased support for fundamental research and education and improved transparency of regulatory processes. This perspectives article summarises the content and conclusions of the workshop and puts forward suggestions on how translational success of cell-based therapies in orthopaedics may be achieved.
Assuntos
Terapia Baseada em Transplante de Células e Tecidos , Ortopedia/tendências , Pesquisa Translacional Biomédica/tendências , Humanos , Engenharia TecidualRESUMO
The "Workshop on Standards & Measurements for Tissue Engineering Scaffolds" was held on May 21, 2013 in Indianapolis, IN, and was sponsored by the ASTM International (ASTM). The purpose of the workshop was to identify the highest priority items for future standards work for scaffolds used in the development and manufacture of tissue engineered medical products (TEMPs). Eighteen speakers and 78 attendees met to assess current scaffold standards and to prioritize needs for future standards. A key finding was that the ASTM TEMPs subcommittees (F04.41-46) have many active "guide" documents for educational purposes, but few standard "test methods" or "practices." Overwhelmingly, the most clearly identified need was standards for measuring the structure of scaffolds, followed by standards for biological characterization, including in vitro testing, animal models and cell-material interactions. The third most pressing need was to develop standards for assessing the mechanical properties of scaffolds. Additional needs included standards for assessing scaffold degradation, clinical outcomes with scaffolds, effects of sterilization on scaffolds, scaffold composition, and drug release from scaffolds. Discussions highlighted the need for additional scaffold reference materials and the need to use them for measurement traceability. Workshop participants emphasized the need to promote the use of standards in scaffold fabrication, characterization, and commercialization. Finally, participants noted that standards would be more broadly accepted if their impact in the TEMPs community could be quantified. Many scaffold standard needs have been identified and focus is turning to generating these standards to support the use of scaffolds in TEMPs.
Assuntos
Engenharia Tecidual/métodos , Engenharia Tecidual/normas , Alicerces Teciduais , Animais , Congressos como Assunto , Educação , HumanosRESUMO
Enhanced tendon and ligament repair would have a major impact on orthopedic surgery outcomes, resulting in reduced repair failures and repeat surgeries, more rapid return to function, and reduced health care costs. Scaffolds have been used for mechanical and biologic reinforcement of repair and regeneration with mixed results. This review summarizes efforts made using biologic and synthetic scaffolds using rotator cuff and ACL as examples of clinical applications, discusses recent advances that have shown promising clinical outcomes, and provides insight into future therapy.
Assuntos
Ligamentos/fisiologia , Regeneração , Tendões/fisiologia , Alicerces Teciduais , Animais , Humanos , Ligamentos/cirurgia , Tendões/cirurgiaRESUMO
There is a recognized and urgent need for improved treatment of articular cartilage defects. Tissue engineering of cartilage using a cell-scaffold approach has demonstrated potential to offer an alternative and effective method for treating articular defects. We have developed a unique, heterogeneous, osteochondral scaffold using the TheriForm three-dimensional printing process. The material composition, porosity, macroarchitecture, and mechanical properties varied throughout the scaffold structure. The upper, cartilage region was 90% porous and composed of D,L-PLGA/L-PLA, with macroscopic staggered channels to facilitate homogenous cell seeding. The lower, cloverleaf-shaped bone portion was 55% porous and consisted of a L-PLGA/TCP composite, designed to maximize bone ingrowth while maintaining critical mechanical properties. The transition region between these two sections contained a gradient of materials and porosity to prevent delamination. Chondrocytes preferentially attached to the cartilage portion of the device, and biochemical and histological analyses showed that cartilage formed during a 6-week in vitro culture period. The tensile strength of the bone region was similar in magnitude to fresh cancellous human bone, suggesting that these scaffolds have desirable mechanical properties for in vivo applications, including full joint replacement.
Assuntos
Materiais Biocompatíveis , Cartilagem Articular/citologia , Cartilagem Articular/transplante , Ácido Láctico/química , Osteoartrite/terapia , Ácido Poliglicólico/química , Polímeros/química , Análise de Variância , Biodegradação Ambiental , Cartilagem Articular/lesões , Colágeno/metabolismo , DNA/metabolismo , Glicosaminoglicanos/química , Humanos , Microscopia Eletrônica de Varredura , Poliésteres , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Sais de Tetrazólio/farmacologia , Tiazóis/farmacologia , CicatrizaçãoRESUMO
This work examines the effect of perfusion on the cell content and sulfated glycosaminoglycan synthesis of ovine articular chondrocytes cultured on polyglycolic acid (PGA) scaffolds. Ovine chondrocytes were seeded onto the scaffolds and cultured for up to 9 days. During this time the cells were subjected to perfusion at velocities of up to 170 microm/s. The samples were radiolabeled with (35)SO(4) to quantify the overall synthesis of sulfated glycosaminoglycans (S-GAGs) and the retention of S-GAGs in the construct. The constructs were also analyzed for DNA as a measure of cellular content. Constructs subjected to perfusion during culture had significantly higher DNA contents than those cultured statically. Matrix metabolism was also modulated by perfusion, with this modulation depending on culture duration. Nine days of continuous perfusion increased S-GAG synthesis and deposition by approximately 40% when compared with static controls. However, perfusion at early time points (during the initial 3-day culture period) suppressed the synthesis and retention of S-GAGs when compared with controls. This work demonstrates the effects of perfusion on cartilage growth in vitro, illustrating the use of perfusion to modulate the growth of tissue-engineered cartilage constructs, and potentially enhance tissue growth in vitro.
Assuntos
Condrócitos/metabolismo , Matriz Extracelular/metabolismo , Engenharia Tecidual , Animais , Técnicas de Cultura de Células/métodos , Perfusão , Ácido Poliglicólico/metabolismo , Ovinos , Engenharia Tecidual/métodosRESUMO
Interactions between bone and cartilage formation are critical during growth and fracture healing and may influence the functional integration of osteochondral repair constructs. In this study, the ability of tissue-engineered cartilage constructs to support bone formation under controlled mechanical loading conditions was evaluated using a lapine hydraulic bone chamber model. Articular chondrocytes were seeded onto polymer disks, cultured for 4 weeks in vitro, and then transferred to empty bone chambers previously implanted into rabbit femoral metaphyses. The effects of chondrocyte viability within the implanted constructs and in vivo mechanical loading on bone formation were tested in separate experiments. After 4 weeks in vivo, biopsies from the chambers consisted of a complex composite of bone, cartilage, and fibrous tissue, with bone forming in direct apposition to the cartilage constructs. Microcomputed tomography imaging of the chamber biopsies revealed that the implantation of viable constructs nearly doubled the bone volume fraction of the chamber tissue from 0.9 to 1.6% as compared with the implantation of devitalized constructs in contralateral control chambers. The application of an intermittent cyclic mechanical load was found to increase the bone volume fraction of the chamber tissue from 0.4 to 3.6% as compared with no-load control biopsies. The results of these experiments demonstrate that tissue-engineered cartilage constructs implanted into a well-vascularized bone defect will support direct appositional bone formation and that bone formation is significantly influenced by the viability of chondrocytes within the constructs and the local mechanical environment in vivo.
Assuntos
Materiais Biocompatíveis , Substitutos Ósseos , Cartilagem , Condrócitos , Engenharia Tecidual , Animais , Imuno-Histoquímica , Masculino , Coelhos , Tomografia Computadorizada por Raios XRESUMO
Bioreactor design in tissue engineering is complex, and at the early stages of its development. Design of biologically effective, yet scalable, devices requires intimate collaboration between engineers and biologists to ensure that all aspects are considered fully. Growth conditions, harvesting time, scale-up, storage, and sterility issues all need to be considered and incorporated into the design of bioreactors. Each tissue-engineered product will likely require individualized bioreactor design. However, without a comprehensive understanding of each of these components, bioreactor design and tissue growth to manufacture product will remain at a relatively rudimentary and limited level. Increased fundamental understanding of the issues can have a dramatic impact on the ability to generate tissue-engineered product safely, economically, and in the numbers that are required to fully address the patient populations in need.
Assuntos
Reatores Biológicos , Técnicas de Cultura de Órgãos/instrumentação , Engenharia Tecidual/instrumentação , Engenharia Tecidual/métodos , Animais , Humanos , Técnicas de Cultura de Órgãos/métodosRESUMO
Static and dynamic compression are known to modulate the metabolism of articular cartilage. The present study focused on determining the effects of compressive loading on the metabolism of sulfated glycosaminoglycans (S-GAG) and protein in tissue engineered cartilage constructs. Cartilage constructs were subjected to static or dynamic compression for 24 h and radiolabeled with 35SO4 and 3H-proline to assess the total synthesis and percentage retention of S-GAG and total protein, respectively. Static compression at an amplitude of 50% suppressed the synthesis of both S-GAG and protein by 35% and 57%, respectively. Dynamic compression at an amplitude of 5% had stimulatory effects on synthesis that were dependent on the static offset compression amplitude (10% or 50%) and dynamic compression frequency (0.001 or 0.1 Hz). Thus, tissue engineered cartilage demonstrated the ability to respond to mechanical loading in a manner similar to that observed with articular cartilage. Mechanical loading may therefore potentially be used to modulate the growth of cartilaginous tissues in vitrd, potentially facilitating the culture of functional cartilage tissues suitable for implantation.
Assuntos
Cartilagem/metabolismo , Engenharia Tecidual , Animais , Fenômenos Biomecânicos , Bovinos , Glicosaminoglicanos/biossíntese , Biossíntese de Proteínas , Estresse MecânicoRESUMO
There are many ways to influence cell activities, and biomaterials with functional groups attached is an attractive method that clearly has the ability to modulate cell behavior. The evidence is clear that biomaterials, with or without growth factors and cells, have resulted in numerous products for the regenerative medicine field. In contrast the functionalized biomaterial products remain in the development phase.
Assuntos
Materiais Biocompatíveis/química , Materiais Biocompatíveis/metabolismo , Medicina Regenerativa/métodos , Matriz Extracelular/química , Humanos , Medicina Regenerativa/instrumentação , Engenharia Tecidual/instrumentação , Engenharia Tecidual/métodosRESUMO
The fields of tissue engineering and regenerative medicine have the capacity to substantially impact clinical care through the introduction of new products that can address unmet clinical needs, or significantly improve on present therapies. These products will be developed through the demonstration of therapeutic effectiveness, adequate safety, and meeting regulatory requirements. The technology used in the product will dictate the product development and manufacturing costs; the regulatory pathway; and the time taken to complete clinical trials, gain regulatory approval, and become commercialized. A comparison of the required investment of time and funds, with the potential revenue generated, allows for a determination of the likely commercialization opportunity. Ultimately, the long-term success of a product will be dependent on its clinical effectiveness and commercial viability.
Assuntos
Osso e Ossos/fisiologia , Comércio , Engenharia Tecidual/economia , Engenharia Tecidual/métodos , Pesquisa Translacional Biomédica/economia , Pesquisa Translacional Biomédica/métodos , Humanos , Investimentos em Saúde , Controle Social Formal , Engenharia Tecidual/legislação & jurisprudência , Pesquisa Translacional Biomédica/legislação & jurisprudênciaRESUMO
The hydraulic resistance R across osteochondral tissue, especially articular cartilage, decreases with degeneration and erosion. Clinically useful measures to quantify and diagnose the extent of cartilage degeneration and efficacy of repair strategies, especially with regard to pressure maintenance, are still developing. The hypothesis of this study was that hydraulic resistance provides a quantitative measure of osteochondral tissue that could be used to evaluate the state of cartilage damage and repair. The aims were to (1) develop a device to measure R in an arthroscopic setting, (2) determine whether the device could detect differences in R for cartilage, an osteochondral defect, and cartilage treated using a hydrogel ex vivo, and (3) determine how quickly such differences could be discerned. The apparent hydraulic resistance of defect samples was ~35% less than intact cartilage controls, while the resistance of hydrogel-filled groups was not statistically different than controls, suggesting some restoration of fluid pressurization in the defect region by the hydrogel. Differences in hydraulic resistance between control and defect groups were apparent after 4 s. The results indicate that the measurement of R is feasible for rapid and quantitative functional assessment of the extent of osteochondral defects and repair. The arthroscopic compatibility of the device demonstrates the potential for this measurement to be made in a clinical setting.
Assuntos
Artroscopia/instrumentação , Cartilagem Articular/lesões , Cartilagem Articular/patologia , Animais , Fenômenos Biomecânicos , Engenharia Biomédica/instrumentação , Doenças das Cartilagens/diagnóstico , Doenças das Cartilagens/cirurgia , Cartilagem Articular/cirurgia , Bovinos , Desenho de Equipamento , Hidrogéis , Técnicas In VitroRESUMO
The National Institutes of Health-sponsored workshop "Translational Models for Musculoskeletal Tissue Engineering and Regenerative Medicine" was held to describe the utility of various translational models for engineered tissues and regenerative medicine therapies targeting intervertebral disc, cartilage, meniscus, ligament, tendon, muscle, and bone. Participants included leaders in the various topics, as well as National Institutes of Health and Food and Drug Administration. The Food and Drug Administration representatives provided perspectives and needs for studies supported by animal models. Researchers described animal models for specific tissues and addressed the following questions: (1) What are the unmet musculoskeletal clinical needs that may be addressed by tissue engineering and regenerative medicine? (2) Are there appropriate models available? (3) Are there needs to develop standardized animal models? (4) What are the translational pathways that lead to clinical trials and therapeutic development? The workshop provided an effective and succinct summary of the status of various animal models in musculoskeletal regenerative medicine. Although many models are available and serve well to answer a variety of questions, the general consensus was that there is a substantial need for improved and standardized animal models for tissue engineering and regenerative medicine of the musculoskeletal system, and that animal models, especially large animal models, are critical to the preclinical step of translating research from bench to bedside.
Assuntos
Modelos Biológicos , Sistema Musculoesquelético , Medicina Regenerativa , Engenharia Tecidual , Pesquisa Translacional Biomédica , Animais , Congressos como Assunto , Modelos Animais de Doenças , Humanos , Meniscos Tibiais/patologia , Camundongos , Ratos , CicatrizaçãoRESUMO
Numerous scaffolds have been proposed for use in connective tissue engineering. Although these scaffolds direct cell migration and attachment, many are biologically inert and thus lack the physiological stimulus to attract cells and induce mitogenesis and matrix synthesis. In the current study, a bioactive scaffold was created by combining a synthetic scaffold with growth factor-rich plasma (GFRP), an autologous concentration of growth factors derived from a platelet-rich plasma preparation. In vitro tendon cell proliferation and matrix synthesis on autologous GFRP-enriched scaffolds, autologous serum-enriched scaffolds, and scaffolds alone were compared. The GFRP preparation was found to have a 4.7-fold greater concentration of a sentinel growth factor (transforming growth factor-beta1) compared with serum. When combined with media containing calcium, the GFRP produced a thin fibrin matrix over and within the GFRP-enriched scaffolds. Cell proliferation assays demonstrated that GFRP-enriched scaffolds significantly enhanced cell proliferation over autologous serum and control groups at both 48 and 72 h. Analysis of the scaffolds at 14, 21, and 28 days revealed that GFRP-enriched scaffolds significantly increased the deposition of a collagen-rich extracellular matrix when compared with the other groups. These results indicate that GFRP can be used to enhance in vitro cellular population and matrix deposition of tissue-engineered scaffolds.