|Year : 2014 | Volume
| Issue : 2 | Page : 155-157
Proof of concept, engineered cartilage tissue for cartilage injuries of knee
K Srinivas Rao
Consultant-Orthopaedics, Employees State Insurance Hospital, Sanathnagar, Hyderabad, Andhra Pradesh, India
|Date of Web Publication||9-Oct-2014|
K Srinivas Rao
Flat no G3, H No. 6-4-519, MIGH Colony, Hyderabad - 500 080, Andhra Pradesh
Source of Support: None, Conflict of Interest: None
Sports related injuries among professional and recreational athletes are increasingly encountered and diagnosed and demand a quick return to preinjury level. At present cartilage injury cases are treated with multiple drilling, abrasion arthroplasty, mosaicplasty, autologous chondrocyte implantation (ACI) and matrix induced autologous chondrocyte implantation (MACI;). The construction of engineered cartilage tissue on a ECM (Extracellular Matrix) composite have high degree of feasibility. Since cartilage cells at the site do not grow to form new cartilage cells,tissue engineered cartilage aim at cultivating chondrocytes in vitro, and to reintroduce the engineered cultured cartilage tissue into the damaged region. To overcome the limitations that currently exist , a multidisciplinary field, in which bioengineering and medicine should emerge).
ان الاصابات الثى تحدثها الرياضة بين الرياضيين المحترفين أوالذين يمارسون الرياض من اجل الترفية تحدث بشكل متزايد ويتم تشخيصها حال جدوثها حتى يتمكن الفرد من العودة السريعة لمستوى لياقته السابقة. وفي وقتنا الحالى ان اصابات الغضروف يتم عالاجها بعدة وسائل مثل الحفر المتعدد، وبواسطة المنظار بالاضافة الى المعالجة بالنقل الذاتى للغضروف، أو النقل الذاتي وزرع خلايا غضروفية . ومن المستجدات فى هذا المجال بناء أنسجة الغضروف هندسيا والتى تبشر بالنجاح. وهناك ايضا الخلايا الغضروفية والتى يمكن انتاجها بطريقة الأنسجة المهندسة في المختبر ومن ثم زراعها فى الموضع الذى اتلف نتيجة الاصابة لتنمو وتشكل خلايا غضروفية جديدة. ولعل نجاح المجهودات الحديثة المشار اليها تمثل حقلا علاجيا جديدا متعدد التخصصات في الهندسة الحيوية والأدوية وغيرها والتى ينبغى التعاون فى ما بينها حتى يثمر وينجح.
Keywords: Extracellular matrix, cartilage, engineering, microcarrier
|How to cite this article:|
Rao K S. Proof of concept, engineered cartilage tissue for cartilage injuries of knee
. Saudi J Sports Med 2014;14:155-7
| Introduction|| |
Cartilage injuries among professional and recreational athletes are increasingly encountered and diagnosed and demand a quick return to preinjury level of sporting activities.  Cartilage lesions in athletically active patients cause considerable morbidity. Especially cartilage injuries of the knee are an increasingly common source of pain and dysfunction, particularly in the athletic population. Current high-field magnetic resonance imaging (MRI) techniques provide a sensitive and reliable diagnostic tool for the evaluation of cartilage and osteochondral injury.  In the athlete, untreated articular cartilage defects can represent a career-threatening injury and create a significant obstacle in returning to full athletic participation. The markedly limited healing potential of articular cartilage often leads to continued deterioration and progressive functional limitations.  Because cartilage cells at the site do not grow to form new cartilage cells, cartilage tissue engineering is required for the repair of injured cartilage. Tissue-engineered cartilage approaches aim at cultivating chondrocytes in vitro and to reintroduce the engineered cultured cartilage tissue into the damaged region. Our proposed cell therapy besides being useful in repairing traumatic cartilage lesions can also serve as bioenhancement procedure after joint surgeries like anterior cruciate ligament (ACL) repair, microfractures, and arthroscopic debridement. Bioenhancement with a bioactive scaffold, like our microcarriercan stimulate healing.  At present, cartilage injury cases are treated with multiple drilling, abrasion arthroplasty, mosaicplasty, autologous chondrocyte implantation (ACI), and matrix-induced ACI (MACI® ). Articular cartilage injuries remain a prime target for regenerative techniques such as tissue engineering. In contrast to the surgical techniques mentioned above, which often lead to the formation of fibrous or fibrocartilaginous tissue, tissue engineering aims at fully restoring the complex structure and properties of the original articular cartilage by using the chondrogenic potential of transplanted cells.  MACI® by Genzyme Corporation is proven for cartilage trauma. Collagen I and collagen III are used for MACI® that provide extracellular matrix (ECM) environment and hence potentially useful.
| Materials and methods|| |
I propose two novel and hypothetical variation of MACI for cartilage injuries. In first approach, autologous chondrocyte are cultured on ECM microcarrier and transplanted. Chondrocyte phenotype can be preserved during culture expansion.  In the second approach, autologous mesenchymal stem cells (MSCs) are cultured on ECM microcarrier and differentiated into hyaline cartilage and used for transplantation. Bone marrow has been shown as a possible source of multipotent stem cells (MSCs) with chondrogenic potential.  MSCs seeded in hydrogel composites can improve cartilage repair.  Effective chondrogenesis can be achieved by directing the MSCs toward chondrocyte lineage.  Both approaches have high degree of feasibility as per the published literature. In the last decades, a wide number of researchers/clinicians involved in tissue engineering field published several works about the possibility to induce a tissue regeneration guided by the use of biomaterials  like our ECM microcarrier. In our study primary chondrocytes were isolated from discarded human cartilage during joint replacement surgeries. The patient's consent was taken for the study. The cartilage cells are isolated after enzyme digestion and are cultured on ECM particle (microcarrier) in spinner bottle set up for required period. Similarly, MSCs isolated from bone marrow concentrates are differentiated into chondrocytes and expanded on ECM microcarrier. The ECM carrier laden with cells is then transplanted in nude mice for study of collagen and GAG expression. In vivo, cartilage matrix formation can be assessed by histology after subcutaneous transplantation of chondrocyte-seeded scaffolds in immunocompromised mice.  The principle of porous microcarrier culture of human or animal cells is described with help of [Figure 1] and [Figure 2].
|Figure 1: Principle of porous microcarrier culture of human cartilage cells|
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| Results|| |
The studies on the construction of engineered cartilage tissue on an ECM composite have high degree of feasibility. The cell attachment ratio to our novel microcarrier, proliferation, and ECM proteins secretion are superior and reproducible. They have appropriate mechanical and structural properties for clinical applications. The scanning electron microscope [Figure 3] and confocal fluoroscope examinations showed that the ECM microcarrier has a regular interconnected porous structure.
|Figure 3: Cartilage cell growing on ECM particle. ECM = Extracellular matrix|
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The novel ECM microcarrier is effective in engineered chondrocyte culture. The cell viability test (WST-1 assay), cell toxicity (lactate dehydrogenase assay), cell survival rate, ECM protein production (glycosaminoglycans contents), cell proliferation (deoxyribonucleic acid (DNA) quantification), and gene expression (real-time polymerase chain reaction (PCR)) all revealed good results for chondrocyte culture. The chondrocytes can maintain normal phenotypes, highly express aggrecan, and type II collagen, and secrete a great deal of ECM when seeded in our novel microcarrier. This study demonstrated that a highly organized "Sol-cell" can be prepared with an ECM microcarrier device that is effective in engineering cartilage tissue.
Some of the in vitro [Figure 3] and in vivo [Figure 4] and [Figure 5] cell culture results are given below here
|Figure 5: Human cartilage cell cultured on ECM and stained with safranin O. ECM = Extracellular matrix|
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| Conclusions|| |
The engineered cartilage tissue will be a promising method for the treatment of cartilage defects. The downsides associated with present treatment regimens like multiple drilling, abrasion arthroplasty, mosaicplasty, and joint replacement can be addressed with our novel, fludic, yet robust cartilage engineering. To overcome the limitations that currently exist, a multidisciplinary field, in which bioengineering and medicine based on integrative approaches using scaffolds, cell populations from different sources, growth factors, and nanomedicine should emerge. 
| References|| |
Kannus P, Natri A. Etiology and pathophysiology of tendon ruptures in sports. Scand J Med Sci Sports 1997;7:107-12.
Gallo RA, Mosher TJ. Imaging of cartilage and osteochondral injuries: A case-based review. Clin Sports Med 2013;32:477-505.
Kane P, Frederick R, Tucker B, Dodson CC, Anderson JA, Ciccotti MG, et al
. Surgical restoration/repair of articular cartilage injuries in athletes. Phys Sportsmed 2013;41:75-86.
Murray MM, Fleming BC. Use of a bioactive scaffold to stimulate anterior cruciate ligament healing also minimizes posttraumatic osteoarthritis after surgery. Am J Sports Med 2013;41:1762-70.
Berninger MT, Wexel G, Rummeny EJ, Imhoff AB, Anton M, Henning TD, et al
. Matrix-assisted autologous chondrocyte transplantation for remodeling and repair of chondral defects in a rabbit model. J Vis Exp 2013;???:e4422.
Heywood HK, Nalesso G, Lee DA, Dell'accio F. Culture expansion in low-glucose conditions preserves chondrocyte differentiation and enhances their subsequent capacity to form cartilage tissue in three-dimensional culture. Biores Open Access 2014;3:9-18.
Gobbi A, Karnatzikos G, Sankineani SR. One-step surgery with multipotent stem cells for the treatment of large full-thickness chondral defects of the knee. Am J Sports Med 2014;42:648-57.
Dashtdar H, Murali MR, Abbas AA, Suhaeb AM, Selvaratnam L, Tay LX, et al
. PVA-chitosan composite hydrogel versus alginate beads as a potential mesenchymal stem cell carrier for the treatment of focal cartilage defects. Knee Surg Sports Traumatol Arthrosc 2013.
Liao J, Hu N, Zhou N, Lin L, Zhao C, Yi S, et al
. Sox9 Potentiates BMP2-Induced chondrogenic differentiation and inhibits BMP2-Induced osteogenic differentiation. PLoS One 2014;9:e89025.
Viti F, Scaglione S, Orro A, Milanesi L. Guidelines for managing data and processes in bone and cartilage tissue engineering. BMC Bioinformatics 2014;15:S14.
Kreuz PC, Gentili C, Samans B, Martinelli D, Krüger JP, Mittelmeier W, et al
. Scaffold-assisted cartilage tissue engineering using infant chondrocytes from human hip cartilage. Osteoarthritis Cartilage 2013;21:1997-2005.
Salgado AJ, Oliveira JM, Martins A, Teixeira FG, Silva NA, Neves NM, et al
. Tissue engineering and regenerative medicine: Past, present, and future. Int Rev Neurobiol 2013;108:1-33.
[Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5]