Content Number: 04
Author Name: Izma Khan
Author I’d: SBPWNC – A04
Educational Institution: Jinnah University For Women, Pakistan
Content Title: Recombinant DNA Technology in the Development of Bio-Materials for Cartilage Regeneration
RECOMBINANT DNA TECHNOLOGY
Recombinant DNA technology was invented by the remarkable work of Stanley N. Cohen, Herbert W. Boyer, and Paul Berg. This technique is primarily used to modify the phenotype of host (organism), when a genetically modified vector incorporates into the genome of an organism
Recombinant DNA has a vast range of use in every aspect of life. It has revolutionized our life by developing such products that offer various opportunities for betterment and innovations that lead to dramatic changes.
Moreover, to gene therapy scientists are moving towards regeneration. It may be of any organ or any tissue.
Regenerative therapy comes up as a remarkable and unique field that consorts biology, robotics, medicine chemistry, computer science, engineering, genetics and many other fields for the acquisition of solutions to medical problems that made the human life and health arduous.
On invasion of any disease or due to any injury, naturally our body has innate immunity that helps our body to heal and defend. What if we help the body to accelerate the healing process? Amalgam approaches can strengthen the body’s natural healing process or can endorse the function of damaged organ.
By words regenerative medicine can be defined as:
“It is the process of regenerating cell, tissues or any organs of human to bring back their original function.”
This powerful field “regenerative therapy” helps to recover structure and functionality of damaged organs and tissues. One of the aims of this therapy is to provide solutions for the organs that are completely damaged.
Advancement in recombinant technology has made it more practicable for doctors to provide expedient cures. For treatment many methods are used that can help the body to heal its very own tissues. One of these techniques is cartilage regeneration. Here we choose to review only cartilage regeneration as there are many strategies and ways to regenerate many tissues and organs.
CARTILAGE
Cartilage is connective tissues with flexibility but less flexible than muscles. It helps to keep the joints movement easy through the coat on bone surfaces.
In adulthood cartilage regenerative ability is zero so if it injured it’s gone permanently and cannot be regrow or repaired. Some surgical procedures can restore cartilage to some extent but cannot bring back to normal condition.
In 1743 Hunter quoted about the cartilage repair
From Hippocrates down to the present age, we shall find that an ulcerating cartilage is found to be a very troublesome disease……and that, when destroyed, it is never recovered.’
Over the previous couple of decades, deoxyribonucleic acid techniques have been verified to be terribly powerful tools for the event of novel protein-based biomaterials that are able to self-assemble into completely different structures, like hydrogels.
There are several ways that may be wont to deliver exogenous cDNAs for the treatment of pathologic or broken gristle. For a fortunate approach, many factors got to be taken into consideration, as well as the extent of gristle pathology, illness processes, and therefore the biological activity of the cistron product. A key element for any cistron medical care application may be a vector that expeditiously delivers the deoxyribonucleic acid (DNA) of interest to the target cell and allows transgene expression of an acceptable level and length to have an effect on the specified biological response. What is more, associate understanding of the natural behavior of the target cell, like its half-life, rate of division, and infectability with the vector are essential to the effectiveness of the procedure. Ways for cistron transfer to osteochondral defects. (a) Vector-seeded artificial matrices. A recombinant vector is adsorbate porous matrix that’s then surgically deep-seated into an osteochondral defect. Bone marrow cells infiltrate the matrix, acquire the transgene and regionally specific chondrogenic factors that stimulate MSCs toward improved repair. (b) Vector seeded bone marrow clot. Contemporary aspirate from the bone marrow is quickly mixed with a recombinant vector, permitting interaction with the cells throughout. The mixture is then allowed to coagulate for 15–30 min. straight off after, the clot containing the genetically changed bone marrow cells is deep-seated into the defect wherever the chondrogenic transgene product area unit regionally synthesized and secreted. Cells gift within the clot further as infiltrating cells area unit then stirred toward improved gristle repair. (c) Cell-seeded bone marrow clot. As an alternative, MSCs is isolated from the bone marrow aspirate, dilated and genetically changed ex vivo, and so seeded into a contemporary aspirate for implantation within the succeeding coagulate.
RECOMBINANT BIOMATERIALS FOR CARTILAGE REGENERATION
Biomaterials with an elastic modulus in the range of 1–10 kPa are of widespread interest, as many native tissues also have moduli in this range. The hydrogels developed to repair joint cartilage are more effective when their stress relaxation behavior matches with the native tissue because such behavior affects load transfer and nutrient transport. Up to 80% of articular cartilage wet weight consists of water. To replicate this environment, hydrogels have become a popular option for cartilage regeneration in situ and cartilage engineering in vitro.
The purpose of these types of scaffolds is not only to provide support for cell attachment and spreading but also to have mechanical stability at the defect site although, it is important to take into account that the aim of these scaffolds is not to substitute for the tissue but to improve cartilage regeneration in order to obtain a mature tissue.
Materials that enhance bone and cartilage regeneration promise to be valuable in both research and clinical applications and the use of recombinant DNA technology has enabled the creation of scaffolds with new levels of bio functionality.
Some of the research that has been done regarding cartilage regeneration is briefly described in the table below.
| Sr. No | Bio-materials | Application of recombinant technology to regenerate cartilage |
| 1 | Recombinant Human Collagen | Collagen is the structural protein that provides support to cartilage and bones. Recombinant collagen has the same sequence of amino acid. These are adequate scaffold for cartilage repair when merge with recombinant bone morphogenetic protein. |
| 2 | Recombinant human type II collagen gel | One of the major cartilage constitute is type II collagen. Bovine chondrocytes isolated and combined with rhCII gels and infuse into mice backs. This technique supports the formation of cartilage. |
| 3 | Bioactive Hydrogel | Delivery of growth factors in cartilage regeneration has great importance but their short half-lives cause problems. For this hindrance bioactive hydrogels are form that can easily mix with cells. |
| 4 | Genetically Engineered Elastin-like Polypeptide | ELPs are manmade polypeptides with repetition of pentapeptide “Val-Pro-Gly-Xaa-Gly”.According to this study chondrocytes that culture in ELP has ability to maintain their round shape morphology by the synthesis of sulfated glycosaminoglycan matrix. This is one of the reasons of great potential in regeneration. |
| 5 | Recombinant glue | For cartilage regeneration recombinant glue is made with the help of Tissue Transglutaminase. |
| 6 | Recombinant Cell Siac | Application of recombinant xeno-free cell saic that are in aggregate-like shape has great ability of regeneration. |
| 7 | Genetically modified rAAV vector- based IGF-1 | rAAV vectors designed by parvoviruses are manufactured from non-pathogenic and non-replicative parvoviruses. Frisch through rAAV vectors with IGF-1 inoculated for medication diseases related to cartilage. |
| 8 | Recombinant muscle and fat grafts | Genetically modified muscles were constructed through adenovirus with BMP-2, Ad luc, Ad GFP and promoters and incorporated into defects of Fischer rats. These muscles show great healing ability within 10 days of transplantation & showed full recovery in 8 weeks. |
| 9 | Recombinant (rhBMP-2) | In adult rabbits 3 mm wide and deep defect created and then filled with rhBMP-2 .After 24 weeks cartilage thickness was 70% of natural cartilage. |
| 10 | Recombinant pNaSS)-Grafted PCL Films | Biocompatible mechanically stable PCL films grafted with pNaSS through recombinant adeno virus that boost process of cartilage. |
COMMERCIALIZATION OF REGENERATIVE MEDICINE FOR CARTILAGE REGENERATION
There are numbers of novel research about cartilage regeneration but the products that are FDA-approved are still limited in number.
Under the section of 351 or 361 of the Public Health Service Act drugs that are used in cartilage regeneration are classified. They are classified as:
- Devices: include scaffolds and injectable drugs
- Drug and Biologics: include stem cell related technologies and biomaterial constructed through protein molecules and growth factors.
Some of the products approved by FDA are mentioned below:
| Sr. No. | Name |
| DEVICE CLASSIFICATION | |
| Devices for cartilage application | |
| 1 | DePuy’s Cartilage Autograft Implantation System (CAIS) |
| 2 | HYTOP from TRB Chemida |
| 3 | Agili C Biphasic Implant |
| 4 | BST-CarGel from Piramal Healthcare |
| Injectible drugs | |
| 1 | Synvisc by Genzyme |
| 2 | Topical NSAID cream Pennsaid by Nuvo Research |
| 3 | Supartz by Smith |
| 4 | Nephew and Mobic by Boehringer Ingelheim |
| 5 | Condrosan by Bioiberica |
| 6 | Dextrose injection by Universidad Nacional de Rosario |
| 7 | TPX-100 |
| BIOLOGICS CLASSIFICATION | |
| 1 | CARTISTEM |
| 2 | TG-C |
| 3 | Revaflex |
| 4 | LNA043 |
CARTILAGE REGENERATION CENTER
The treatment of cartilage injuries remains one of the most difficult challenges in medicine. Name of some renowned center are listed below:
| S.no | Names |
| 1. | Meniscus Tear-Agha Khan University Hospital |
| 2. | CRC Brigham and Women’s Hospital |
| 3. | UCSF Orthopaedic Institute |
| 4. | CRC at Bone & Joint Clinic of Baton Rouge |
| 5. | Shifa Regenerative & Rejuvenation Clinic |
| 6. | Health Care Cartilage Repair & Restoration |
| 7. | Paley Orthopedic & Spine Institute |
| 8. | Center for Advanced Cartilage Repair and Osteochondritis Dissecans Treatment |
FUTURE PROSPECTS
Cartilage regeneration takes 20 years to achieve dignitary place, and current treatments try to make it possible that modified cartilage tissues to come close to natural ones. It should be kept in mind that biotechnology is an interdisciplinary subject that provides new techniques and concepts. Related to cartilage regeneration, modern trend deals with the potential of biotechnology in the textile industry. It broadens the utilization of present-day fabrics. Fabrics are made from innovative materials that have the ability to regenerate cartilage. This technique would have many beneficial outcomes for patients. Patients would not undergo surgical treatment or any specific therapy they can use in their daily life without any hesitation, moreover it does not cause morbidity. Advancement in material designing will boost our capability to reconstruct cartilage with great functionality. Although there are some challenges in a way, research overcomes those challenges through their novel ideas and hope for betterment in the field of cartilage regeneration.
REFERENCES
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