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1.
Int J Mol Sci ; 22(19)2021 Sep 22.
Article in English | MEDLINE | ID: mdl-34638538

ABSTRACT

Osteoarthritis (OA) is a chronic debilitating disorder causing pain and gradual degeneration of weight-bearing joints with detrimental effects on cartilage volume as well as cartilage damage, generating inflammation in the joint structure. The etiology of OA is multifactorial. Currently, therapies are mainly addressing the physical and occupational aspects of osteoarthritis using pharmacologic pain treatment and/or surgery to manage the symptomatology of the disease with no specific regard to disease progression or prevention. Herein, we highlight alternative therapeutics for OA specifically considering innovative and encouraging translational methods with the use of adipose mesenchymal stem cells.


Subject(s)
Adipose Tissue/cytology , Chondrocytes/transplantation , Mesenchymal Stem Cell Transplantation/methods , Osteoarthritis, Knee/therapy , Regenerative Medicine/methods , Cartilage, Articular/pathology , Humans , Mesenchymal Stem Cells/cytology , Osteoarthritis, Knee/pathology , Stress, Mechanical
2.
Tissue Eng ; 13(11): 2733-41, 2007 Nov.
Article in English | MEDLINE | ID: mdl-17880270

ABSTRACT

Keratinocyte stem cell technology provides at least an adjuvant therapy to clinically close large cutaneous wounds (e.g., burn wounds). Here, the performance of keratinocyte cultures depends primarily on the quality of the bed to which they are applied. Clinical take rates for cultured keratinocyte grafts are optimal when applied to a vascularized dermal bed with minimal bacterial colonization. In the absence of autologous dermis, staged reconstruction with a dermal equivalent or dermal regeneration template is required. A novel product, Hyalomatrix, is a bilayer of an esterified hyaluronan scaffold beneath a silicone membrane. The scaffold delivers hyaluronan to the wound bed, and the silicone membrane acts as a temporary epidermal barrier. The product has been investigated in a controlled, porcine, acute full-thickness excisional wound model. Cultured autologous keratinocytes (CAKs) were delivered on Laserskin to acute full-thickness wounds treated with Hyalomatrix within chambers, and graft take rates were assessed longitudinally using image analysis. In the absence of chambers, wound contraction was assessed. Clinical CAK take rates fall sequentially with delay in application post-Hyalomatrix pre-treatment, but repeated pre-treatment removed this, with maximal take of 57.2% at 5 weeks post-wounding. In the absence of chambers, more-complete wound closure resulted from edge re-epithelialization and contraction, by a factor of 5 at 1 month, and was achieved at least 2 weeks sooner in the gold standard controls of split-thickness autograft to an acute or pre-treated wound bed. Wound contraction and late neodermal morphology (1 year) were similar in pre-treated CAKs and split-thickness autograft wounds. In this model, the Hyalomatrix wound bed pre-treatment increase in CAK take appeared to be dose dependent. The product appeared to act as a hyaluronan delivery system rather than a dermal regeneration template. The silicone membrane may limit wound bed colonization, and the combination of this temporary barrier with hyaluronan delivery and neodermis induction has been termed a barrier-delivery-induction system. The development of similar systems for serial application offers an alternative to a dermal regeneration template when CAKs are engrafted in the hostile, colonized environment of large burn wounds.


Subject(s)
Hyaluronic Acid , Keratinocytes/transplantation , Skin, Artificial , Stem Cells/cytology , Wound Healing , Animals , Cells, Cultured , Disease Models, Animal , Epidermal Cells , Male , Skin Transplantation , Swine , Swine, Miniature , Time Factors , Transplantation, Autologous , Treatment Outcome
3.
Tissue Eng ; 12(7): 1787-98, 2006 Jul.
Article in English | MEDLINE | ID: mdl-16889509

ABSTRACT

The regeneration of damaged organs requires that engineered tissues mature when implanted at sites of injury or disease. We have used new analytic techniques to determine the extent of tissue regeneration after treatment of knee injury patients with a novel cartilage tissue engineering therapy and the effect of pre-existing osteoarthritis on the regeneration process. We treated 23 patients, with a mean age of 35.6 years, presenting with knee articular cartilage defects 1.5 cm2 to 11.25 cm2 (mean, 5.0 cm2) in area. Nine of the patients had X-ray evidence of osteoarthritis. Chondrocytes were isolated from healthy cartilage removed at arthroscopy. The cells were cultured for 14 days, seeded onto esterified hyaluronic acid scaffolds (Hyalograft C), and grown for a further 14 days before implantation. A second-look biopsy was taken from each patient after 6 to 30 months (mean, 16 months). After standard histological analysis, uncut tissue was further analyzed using a newly developed biochemical protocol involving digestion with trypsin and specific, quantitative assays for type II collagen, type I collagen, and proteoglycan, as well as mature and immature collagen crosslinks. Cartilage regeneration was observed as early as 11 months after implantation and in 10 out of 23 patients. Tissue regeneration was found even when implants were placed in joints that had already progressed to osteoarthrosis. Cartilage injuries can be effectively repaired using tissue engineering, and osteoarthritis does not inhibit the regeneration process.


Subject(s)
Bioprosthesis , Cartilage/transplantation , Chondrocytes/transplantation , Hyaluronic Acid , Osteoarthritis, Knee/therapy , Regeneration , Tissue Engineering , Adolescent , Adult , Cartilage/metabolism , Chondrocytes/metabolism , Extracellular Matrix Proteins/biosynthesis , Female , Humans , Male , Middle Aged , Time Factors
4.
Tissue Eng ; 11(1-2): 277-87, 2005.
Article in English | MEDLINE | ID: mdl-15758586

ABSTRACT

Reliable and reproducible outcome measures are essential to assess the efficacy of competing and novel tissue-engineering techniques. The aim of this study was to compare traditional histological analyses with newly developed quantitative biochemical outcome measures for the repair of articular cartilage. The production of a new anti-peptide antibody and the development and validation of a novel method for the extraction and immunoassay of type I collagen are described. The assay was used, in conjunction with existing assays for type II collagen and proteoglycans, to measure levels of the matrix components in repair tissue biopsies obtained from patients treated with the new tissue-engineering therapy Hyalograft C. Frozen sections cut from the same biopsies were stained for proteoglycans, using safranin O, and immunohistochemical analysis was used to assess type I and II collagen staining. Although there was general agreement between the extent of staining and the amounts of the three matrix components, there was a large degree of overlap in biochemical content between biopsies classified histologically on the basis of low, moderate, or abundant staining. The results demonstrate that histological grading of matrix protein abundance to classify repair cartilage as hyaline or fibrocartilagenous is often misleading. In addition, we demonstrate for the first time the ability to measure collagen cross-links in repair tissue biopsies and show that it can be used as a surrogate marker for tissue maturity. Our new range of biochemical techniques provides a standardized method to assess the quality of both engineered cartilage produced in vitro and repair tissue biopsies obtained after in vivo implantation.


Subject(s)
Cartilage Diseases/drug therapy , Cartilage, Articular , Chondrocytes/metabolism , Tissue Engineering/methods , Animals , Biopsy , Cartilage Diseases/surgery , Cattle , Cells, Cultured , Chondrocytes/cytology , Collagen Type I/analysis , Collagen Type I/drug effects , Collagen Type II/analysis , Collagen Type II/drug effects , Enzyme-Linked Immunosorbent Assay , Frozen Sections , Humans , Immunohistochemistry , Proteoglycans/metabolism , Rats , Reproducibility of Results , Second-Look Surgery , Treatment Outcome , Trypsin/pharmacology
6.
J Periodontol ; 74(2): 262-7, 2003 Feb.
Article in English | MEDLINE | ID: mdl-12666716

ABSTRACT

BACKGROUND: An autologous cell hyaluronic acid graft was used for gingival augmentation in mucogingival surgery. METHODS: Seven sites from 6 patients were used in this study. Five patients (5 sites) needed gingival augmentation prior to prosthetic rehabilitation, and one patient (2 sites) needed augmentation because of pain during daily toothbrushing. Full-mouth plaque score (FMPS), full-mouth bleeding score (FMBS), probing depth (PD), and clinical attachment level (CAL) were recorded for the sites at baseline and 3 months after surgery. The amount of keratinized tissue (KT) was measured in the mesial, middle, and distal sites of each involved tooth. A small 2 x 1 x 1 mm portion of gingiva (epithelium and connective tissue) was removed from each patient, placed in a nutritional medium, and sent to the laboratory. The gingival tissue was processed: keratinocytes and fibroblasts were separated and only fibroblasts were cultivated. They were cultured on a scaffold of fully esterified benzyl ester hyaluronic acid (HA) and returned to the periodontal office under sterile conditions. During the gingival augmentation procedure, the periosteum of the selected teeth was exposed, and the membrane containing cultivated fibroblasts was adapted to and positioned on the site. RESULTS: Three months after surgery, an increased amount of gingiva was obtained, and the histological examination revealed a fully keratinized tissue on all the treated sites. CONCLUSION: Tissue engineering technology using an autologous cell hyaluronic acid graft was applied in gingival augmentation procedures and provides an increase of gingiva in a very short time without any discomfort for the patient.


Subject(s)
Cell Culture Techniques , Fibroblasts/transplantation , Hyaluronic Acid , Membranes, Artificial , Tissue Engineering/methods , Vestibuloplasty/methods , Absorbable Implants , Adult , Female , Gingiva/physiology , Gingivoplasty/methods , Humans , Male , Regeneration
7.
Novartis Found Symp ; 249: 218-29; discussion 229-33, 234-8, 239-41, 2003.
Article in English | MEDLINE | ID: mdl-12708659

ABSTRACT

Outcome measures for cartilage repair techniques include clinical assessment of functional status, magnetic resonance imaging, mechanical indentation in situ and second-look biopsies, which are used for detailed ex vivo histological and immunohistochemical assessment. Biopsy analysis is considered an important outcome measure, despite being highly invasive, since it provides a visual record of the spatial organization of matrix proteins and cells. We propose that the value of second-look biopsies would be significantly enhanced if accurate quantification of cartilage matrix molecules could also be obtained. The goal of our work has been to develop a combined method for histological and biochemical analysis of a single biopsy. We have developed a method of cutting frozen sections of cartilage and recovering the uncut tissue for subsequent biochemical analysis. We have also developed a range of miniaturized assays that can be performed after cartilage digestion with trypsin. In this way we are now able to analyse biopsies with a wet weight as low as 5 mg using both histological and biochemical methods, so obtaining the maximum amount of information from the minimum volume of tissue. This new approach will allow a more accurate assessment of the quality of cartilage repair tissue than histological analysis alone.


Subject(s)
Biocompatible Materials , Cartilage, Articular/pathology , Chondrocytes/transplantation , Hyaluronic Acid , Prostheses and Implants , Surgical Mesh , Tissue Engineering , Age Factors , Animals , Biopsy , Cartilage, Articular/chemistry , Cartilage, Articular/injuries , Cartilage, Articular/surgery , Cattle , Cells, Cultured/chemistry , Cells, Cultured/transplantation , Chondrocytes/chemistry , Collagen Type I/analysis , Collagen Type II/analysis , Endopeptidases/pharmacology , Enzyme-Linked Immunosorbent Assay , Follow-Up Studies , Frozen Sections , Glycosaminoglycans/analysis , Hyalin/chemistry , Hydroxyproline/analysis , Organ Size , Rabbits , Second-Look Surgery
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