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      Viability and Regeneration of Chondrocytes after Laser Cartilage Reshaping Using 1,460 nm Diode Laser

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          Abstract

          Objectives

          Cartilage reshaping by laser irradiation is used to correct septal and auricular cartilage deformities. Chondrocyte viability following laser irradiation and reshaping has been well established. However, the regeneration process of chondrocyte after laser irradiation has not been revealed yet. The aims of this study were to determine the mechanism of cartilaginous thermal injury and the regenerative process of damaged cartilage following laser irradiation.

          Methods

          Laser irradiation was performed on human septal cartilage and rabbit auricular cartilage using a 1,460-nm diode laser. We observed change in the shape of cartilage and evaluated the extent of cartilage injury using live/dead cell assay via confocal microscopy. Hoechst and propidium iodide (PI) staining was used to evaluate the mechanism of chondrocyte injury after laser irradiation. To evaluate the regeneration of cartilage, laser irradiated cartilages were reimplanted into a subperichondrial pocket and were harvested at 1, 2, and 4 weeks after reimplantation for viability assessment and histologic examination.

          Results

          Laser irradiation using a 1,460-nm diode laser produced a marked shape change in both human septal and rabbit auricular cartilages. Thermal damage on cartilage was correlated with the exposure time and the laser power. Hoechst and PI staining showed that chondrocyte death by laser irradiation was due to mainly necrosis, rather than apoptosis. In lower power treatment group (0.3 W and 0.5 W), all the chondrocytes regenerated within 4 weeks, however, in 1 W treatment group, chondrocytes could not regenerate until 4 weeks.

          Conclusion

          Reshaping of cartilage using 1,460 nm diode laser was attained concurrently with the thermal injury to the chondrocytes. The extent of thermal damage on chondrocytes was dependent on the exposure time and the laser power and the damaged chondrocytes irradiated with lower level of laser power could be regenerated after reimplantation into subperichondrial pocket.

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          The effect of monopolar radiofrequency energy on partial-thickness defects of articular cartilage.

          Y Lu, K Hayashi, P Hecht … (2015)
          To evaluate the effect of monopolar radiofrequency (RF) energy on partial-thickness defects of articular cartilage, comparing the outcome of partial-thickness defects treated with monopolar RF energy with that of treatment by conversion of partial-thickness defects to full-thickness defects by curettage and microfracture. Randomized trial using adult female sheep. Thirty-six sheep were used in this study. Both stifles in each animal were randomly assigned to 1 of the following 3 procedures: (1) partial-thickness defect without any treatment to serve as a sham-operated control, (2) partial-thickness defect with RF energy treatment, and (3) partial-thickness defect treated by conversion of the defect to a full-thickness defect by curettage and microfracture. Nine sheep were euthanized at 0, 2, 12, and 24 weeks after surgery (n = 6 per group). After euthanasia, cartilage samples were harvested from the defect sites, and chondrocyte viability was analyzed by confocal laser microscopy using a triple-labeling technique. Cartilage samples also were decalcified and stained with hematoxylin and eosin and safranin-O for histologic analysis. Surface properties of cartilage samples were analyzed using scanning electron microscopy. The analysis of chondrocyte viability showed that RF treatment caused death of almost all chondrocytes in the defect. Histologic analysis showed that RF treatment caused detrimental effects to chondrocytes and proteoglycan concentration that progressed over time, and that full-thickness defects were repaired by fibrocartilage by 24 weeks after surgery. Scanning electron microscopy analysis indicated that RF-treated groups were significantly smoother and less irregular than control groups at 2, 12, and 24 weeks after surgery. This study showed that monopolar RF energy caused long-term damage to cartilage in this sheep model and did not appear to have the beneficial effects reported in a previous study that evaluated application of this technique using a bipolar RF probe.
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            The dual role of perichondrium in cartilage wound healing.

            Mark Duynstee, Henriette L. Verwoerd-Verhoef, Dania van der Meer … (2002)
            Cartilage structures from the head and neck possess a certain but limited capacity to heal after injury. This capacity is accredited to the perichondrium. In this study, the role of the inner (cambium) and the outer (fibrous) layers of the perichondrium in cartilage wound healing in vitro is investigated. For the first time, the possibility of selectively removing the outer perichondrium layer is presented. Using rabbit ears, three different conditions were created: cartilage explants with both perichondrium layers intact, cartilage explants with only the outer perichondrium layer dissected, and cartilage explants with both perichondrium layers removed. The explants were studied after 0, 3, 7, 14, and 21 days of in vitro culturing using histochemistry and immunohistochemistry for Ki-67, collagen type II, transforming growth factor beta 1 (TGFbeta1), and fibroblast growth factor 2 (FGF2). When both perichondrium layers were not disturbed, fibrous cells grew over the cut edges of the explants from day 3 of culture on. New cartilage formation was never observed in this condition. When only the outer perichondrium layer was dissected from the cartilage explants, new cartilage formation was observed around the whole explant at day 21. When both perichondrium layers were removed, no alterations were observed at the wound surfaces. The growth factors TGFbeta1 and FGF2 were expressed in the entire perichondrium immediately after explantation. The expression gradually decreased with time in culture. However, the expression of TGFbeta1 remained high in the outer perichondrium layer and the layer of cells growing over the explant. This indicates a role for TGFbeta1 in the enhancement of fibrous overgrowth during the cartilage wound-healing process. The results of this experimental in vitro study demonstrate the dual role of perichondrium in cartilage wound healing. On the one hand, the inner layer of the perichondrium, adjacent to the cartilage, provides (in time) cells for new cartilage formation. On the other hand, the outer layer rapidly produces fibrous overgrowth, preventing the good cartilage-to-cartilage connection necessary to restore the mechanical function of the structure.
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              Effect of bipolar radiofrequency energy on human articular cartilage. Comparison of confocal laser microscopy and light microscopy.

              B. Cole, Mark D. Markel, R Edwards … (2001)
              To evaluate chondrocyte viability using confocal laser microscopy (CLM) following exposure to bipolar radiofrequency energy (bRFE) and to contrast CLM with standard light microscopy (LM) techniques. In vitro analysis using chondromalacic human cartilage. Twelve fresh chondral specimens were treated with the ArthroCare 2000 bRFE system (ArthroCare, Sunnyvale, CA) coupled with 1 of 2 types of probes and at 3 energy delivery settings (S2, S4, S6). A sham-operated group was treated with no energy delivered. Specimens were analyzed for chondrocyte viability and chondral morphology with CLM using fluorescent vital cell staining and with LM using H&E and safranin-O staining. LM with H&E staining showed smoothing of fine fronds of fibrillated cartilage; thickened fronds were minimally modified. Chondrocyte nuclei were present and not morphologically different than nuclei within sham-operated and adjacent untreated regions. LM with safranin-O staining showed a clear demarcation between treated and untreated regions. CLM, however, showed chondrocyte death: the depth and width of chondrocyte death increased with increasing bRFE settings. CLM showed that bRFE delivered through the probes investigated created significant chondrocyte death. These changes were not apparent using LM techniques.
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                Author and article information

                Journal
                Journal ID (nlm-ta): Clin Exp Otorhinolaryngol
                Journal ID (iso-abbrev): Clin Exp Otorhinolaryngol
                Journal ID (publisher-id): CEO
                Title: Clinical and Experimental Otorhinolaryngology
                Publisher: Korean Society of Otorhinolaryngology-Head and Neck Surgery
                ISSN (Print): 1976-8710
                ISSN (Electronic): 2005-0720
                Publication date (Print): June 2013
                Publication date (Electronic): 14 June 2013
                Volume: 6
                Issue: 2
                Pages: 82-89
                Affiliations
                [1 ]Department of Otorhinolaryngology, Dankook University College of Medicine, Cheonan, Korea.
                [2 ]Beckman Laser Institute-Korea, Dankook University, Cheonan, Korea.
                [3 ]Department of Otorhinolaryngology, Eulji Hospital, Seoul, Korea.
                Author notes
                Corresponding author: Young-Jun Chung. Department of Otorhinolaryngology, Dankook University College of Medicine, 119 Dandae-ro, Dongnam-gu, Cheonan 330-714, Korea. Tel: +82-41-550-3974, Fax: +82-41-556-1090, docjung@ 123456paran.com

                *These authors contributed equally to this paper.

                Article
                DOI: 10.3342/ceo.2013.6.2.82
                PMC ID: 3687067
                PubMed ID: 23799165
                SO-VID: 691bc423-36bd-43e5-a244-3a156b01474c
                Copyright © Copyright © 2013 by Korean Society of Otorhinolaryngology-Head and Neck Surgery.
                License:

                This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                Date received : 02 April 2013
                Date revision received : 01 May 2013
                Date accepted : 02 May 2013
                Categories
                Subject: Original Article

                ScienceOpen disciplines: Otolaryngology
                Keywords: laser,reshaping,diode laser,chondrocyte,cartilage,septoplasty
                Data availability:
                ScienceOpen disciplines: Otolaryngology
                Keywords: laser, reshaping, diode laser, chondrocyte, cartilage, septoplasty

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