Human Nasal Septal Chondrocytes Research Articles

Effects of Chitosan on Chondrocytes Derived from Human Nasal Septal Cartilage.

The aim of this study was to evaluate the influence of chitosan on the growth of nasal septal chondrocytes (NSCs). The final goal was to establish a novel methodology to enhance nasal septal cartilage regeneration. Human NSCs were isolated and their morphology was examined using Alcian blue staining and observed by light microscopy. The isolated NSCs were grown with various concentrations of chitosan and the expression of COL2A1 was investigated. NSCs were successfully isolated from nasal septal cartilage. Co-culture with 0.2% of chitosan greatly enhanced proliferation of NSCs compared to control cells. However, 0.5% of chitosan was harmful to NSCs, resulting in cell detachment from the culture plate. Furthermore, the addition of 0.2% chitosan significantly improved the expression of COL2A1 in NSCs. To our knowledge, this is the first report to demonstrate that chitosan could effectively guide the attachment and growth of human NSCs. Chitosan appears to be a promising additive for NSC culture, which sets the stage for studying tissue regeneration in nasal septal cartilage deficiency, rhinoplasty, and craniofacial reconstruction.

Human nasal septal chondrocytes (NSCs) preconditioned on NSC-derived matrix improve their chondrogenic potential

BackgroundExtracellular matrix (ECM) has a profound effect on cell behaviors. In this study, we prepare a decellularized human nasal septal chondrocyte (NSC)-derived ECM (CHDM), as a natural (N-CHDM) or soluble form (S-CHDM), and investigate their impact on NSCs differentiation.MethodsN-CHDM, S-CHDM were obtained from NSC. To evaluate function of NSC cultured on each substrate, gene expression using chondrogenic marker, and chondrogenic protein expression were tested. Preconditioned NSCs-loaded scaffolds were transplanted in nude mice for 3 weeks and analyzed.ResultsWhen cultivated on each substrate, NSCs exhibited similar cell spread area but showed distinct morphology on N-CHDM with significantly lower cell circularity. They were highly proliferative on N-CHDM than S-CHDM and tissue culture plastic (TCP), and showed more improved cell differentiation, as assessed via chondrogenic marker (Col2, Sox9, and Aggrecan) expression and immunofluorescence of COL II. We also investigated the effect of NSCs preconditioning on three different 2D substrates while NSCs were isolated from those substrates, subsequently transferred to 3D mesh scaffold, then cultivated them in vitro or transplanted in vivo. The number of cells in the scaffolds was similar to each other at 5 days but cell differentiation was notably better with NSCs preconditioned on N-CHDM, as assessed via real-time q-PCR, Western blot, and immunofluorescence. Moreover, when those NSCs-loaded polymer scaffolds were transplanted subcutaneously in nude mice for 3 weeks and analyzed, the NSCs preconditioned on the N-CHDM showed significantly advanced cell retention in the scaffold, more cells with a chondrocyte lacunae structure, and larger production of cartilage ECM (COL II, glycosaminoglycan).ConclusionsTaken together, a natural form of decellularized ECM, N-CHDM would present an advanced chondrogenic potential over a reformulated ECM (S-CHDM) or TCP substrate, suggesting that N-CHDM may hold more diverse signaling cues, not just limited to ECM component.

Histological Image Processing for the Assessment of Tissue Engineered Cartilage

Abstract In a recently developed perfusion bioreactor, human nasal septum chondrocytes were seeded on decellularized porcine collagen scaffolds and cultivated for 6 weeks. Afterwards, the samples were stained with alcian blue and multiple histological microscope images were taken to assess the migration of the human cells and their redifferentiation. In this paper, we propose an image processing algorithm to objectively quantify the cultivation results. This algorithm performs a white balance, detects cell nuclei and their migration depth, and detects lacunae and their orientation. To evaluate performance, sensitivity and precision are calculated using manually labeled data over 50 images. The detection of cell nuclei has a reasonable precision (73%), but the level of sensitivity (42%) should be further improved. The proposed method demonstrates acceptable level of precision (83%) and sensitivity (86%) for the detection of lacunae.

Laser surface modification of decellularized extracellular cartilage matrix for cartilage tissue engineering.

The implantation of autologous cartilage as the gold standard operative procedure for the reconstruction of cartilage defects in the head and neck region unfortunately implicates a variety of negative effects at the donor site. Tissue-engineered cartilage appears to be a promising alternative. However, due to the complex requirements, the optimal material is yet to be determined. As demonstrated previously, decellularized porcine cartilage (DECM) might be a good option to engineer vital cartilage. As the dense structure of DECM limits cellular infiltration, we investigated surface modifications of the scaffolds by carbon dioxide (CO2) and Er:YAG laser application to facilitate the migration of chondrocytes inside the scaffold. After laser treatment, the scaffolds were seeded with human nasal septal chondrocytes and analyzed with respect to cell migration and formation of new extracellular matrix proteins. Histology, immunohistochemistry, SEM, and TEM examination revealed an increase of the scaffolds' surface area with proliferation of cell numbers on the scaffolds for both laser types. The lack of cytotoxic effects was demonstrated by standard cytotoxicity testing. However, a thermal denaturation area seemed to hinder the migration of the chondrocytes inside the scaffolds, even more so after CO2 laser treatment. Therefore, the Er:YAG laser seemed to be better suitable. Further modifications of the laser adjustments or the use of alternative laser systems might be advantageous for surface enlargement and to facilitate migration of chondrocytes into the scaffold in one step.

THE EFFECTS OF PALMVITEE ON HUMAN NASAL SEPTAL CHONDROCYTES CULTURE EXPANSION AND CARTILAGE RECONSTRUCTION

Palmvitee may act as a beneficial supplement to cartilage tissue engineering which is currently still has certain limitation on chondrocyte expansion and dedifferentiation. Our objectives in thisstudy were to evaluate the effects of Palmvitee on the growth kinetic and phenotype gene expression of human nasal septal chondrocytesin monolayer expansion as well as cartilage reconstruction via tissue engineering technology. Human chondrocytes were cultured inmediums containing various Palmvitee concentrations. Among the different test groups, the medium containing Palmvitee at 3 μg/ml supported the highest chondrocyte growth rate. The gene expression in monolayer chondrocytes culture supplemented with 3 μg/ml Palmviteedemonstrated similar results as in control. Cultured chondrocytes from the medium added with 3 μg/ml Palmvitee were then mixed with Pluronic F127 for cartilage reconstruction in nude mice model. Palmvitee supplementation supported the engineered cartilage development as shown in the histological and gene expression analysis on engineered tissue after 8 weeks of implantation. Therefore, Palmvitee at 3 μg/ml is beneficial for the human nasal septal chondrocytes monolayer expansion and cartilage engineering.

Automated bioreactor system for cartilage tissue engineering of human primary nasal septal chondrocytes.

An automated bioreactor system for three-dimensional (3D) cultivation of facial cartilage replacement matrices (e.g. whole human auricles) with automatised medium exchange, gas flow and temperature control was developed. The measurement of O2 saturation and pH value in the medium was performed with a non-invasive optical method. The whole system can be observed via remote monitoring worldwide. First results demonstrated that the complete system remained sterile throughout a period of 42 days. Human chondrocytes migrated into the employed cartilage replacement matrix consisting of decellularised porcine nasoseptal cartilage (pNSC). Furthermore, an improved migration and new synthesis of aggrecan was detected. A first evaluation of the system was conducted by comparison of the results from laboratory analysis with computational fluid dynamics (CFD).

New bioreactor vessel for tissue engineering of human nasal septal chondrocytes

AbstractCultivation of human nasal septal chondrocytes in a self-established automated bioreactor system with a new designed reactor glass vessel and the results of a computational fluid dynamics model are presented. The first results show the effect of a homogeneous fluidic condition of the continuous medium flow and the resulting stresses on the scaffolds’ surface and their influence on the migration of the cells into the scaffold matrix under these conditions. For this purpose computational models, generated with the computational fluid dynamics software STAR-CCM+, and the results of alcian blue staining for newly synthesized sulphated glycosaminoglycans have been compared during cultivation in the new and a first version of the glass reactor vessel with inhomogeneous fluidic conditions, with the same automated bioreactor system and under similar cultivation conditions.

Characterization of a migrative subpopulation of adult human nasoseptal chondrocytes with progenitor cell features and their potential for in vivo cartilage regeneration strategies.

BackgroundProgenitor cells display interesting features for tissue repair and reconstruction. In the last years, such cells have been identified in different cartilage types. In this study, we isolated a migrative subpopulation of adult human nasoseptal chondrocytes with progenitor cell features by outgrowth from human nasal septum cartilage. These putative progenitor cells were comparatively characterized with mesenchymal stem cells (MSC) and human nasal septum chondrocytes with respect to their cellular characteristics as well as surface marker profile using flow cytometric analyses. Differentiation capacity was evaluated on protein and gene expression levels.ResultsThe migrative subpopulation differentiated into osteogenic and chondrogenic lineages with distinct differences to chondrocytes and MSC. Cells of the migrative subpopulation showed an intermediate surface marker profile positioned between MSC and chondrocytes. Significant differences were found for CD9, CD29, CD44, CD90, CD105 and CD106. The cells possessed a high migratory ability in a Boyden chamber assay and responded to chemotactic stimulation. To evaluate their potential use in tissue engineering applications, a decellularized septal cartilage matrix was either seeded with cells from the migrative subpopulation or chondrocytes. Matrix production was demonstrated immunohistochemically and verified on gene expression level. Along with secretion of matrix metalloproteinases, cells of the migrative subpopulation migrated faster into the collagen matrix than chondrocytes, while synthesis of cartilage specific matrix was comparable.ConclusionsCells of the migrative subpopulation, due to their migratory characteristics, are a potential cell source for in vivo regeneration of nasal cartilage. The in vivo mobilization of nasal cartilage progenitor cells is envisioned to be the basis for in situ tissue engineering procedures, aiming at the use of unseeded biomaterials which are able to recruit local progenitor cells for cartilage regeneration.

Effect of oxygen tension on tissue-engineered human nasal septal chondrocytes.

Tissue-engineered nasal septal cartilage may provide a source of autologous tissue for repair of craniofacial defects. Although advances have been made in manipulating the chondrocyte culture environment for production of neocartilage, consensus on the best oxygen tension for in vitro growth of tissue-engineered cartilage has not been reached. The objective of this study was to determine whether in vitro oxygen tension influences chondrocyte expansion and redifferentiation. Proliferation of chondrocytes from 12 patients expanded in monolayer under hypoxic (5% or 10%) or normoxic (21%) oxygen tension was compared over 14 days of culture. The highest performing oxygen level was used for further expansion of the monolayer cultures. At confluency, chondrocytes were redifferentiated by encapsulation in alginate beads and cultured for 14 days under hypoxic (5 or 10%) or normoxic (21%) oxygen tension. Biochemical and histological properties were evaluated. Chondrocyte proliferation in monolayer and redifferentiation in alginate beads were supported by all oxygen tensions tested. Chondrocytes in monolayer culture had increased proliferation at normoxic oxygen tension (p = 0.06), as well as greater accumulation of glycosaminoglycan (GAG) during chondrocyte redifferentiation (p < 0.05). Chondrocytes released from beads cultured under all three oxygen levels showed robust accumulation of GAG and type II collagen with a lower degree of type I collagen immunoreactivity. Finally, formation of chondrocyte clusters was associated with decreasing oxygen tension (p < 0.05). Expansion of human septal chondrocytes in monolayer culture was greatest at normoxic oxygen tension. Both normoxic and hypoxic culture of human septal chondrocytes embedded in alginate beads supported robust extracellular matrix deposition. However, GAG accumulation was significantly enhanced under normoxic culture conditions. Chondrocyte cluster formation was associated with hypoxic oxygen tension.

Marine Collagen Scaffolds for Nasal Cartilage Repair: Prevention of Nasal Septal Perforations in a New Orthotopic Rat Model Using Tissue Engineering Techniques

Autologous grafts are frequently needed for nasal septum reconstruction. Because they are only available in limited amounts, there is a need for new cartilage replacement strategies. Tissue engineering based on the use of autologous chondrocytes and resorbable matrices might be a suitable option. So far, an optimal material for nasal septum reconstruction has not been identified. The aim of our study was to provide the first evaluation of marine collagen for use in nasal cartilage repair. First, we studied the suitability of marine collagen as a cartilage replacement matrix in the context of in vitro three dimensional cultures by analyzing cell migration, cytotoxicity, and extracellular matrix formation using human and rat nasal septal chondrocytes. Second, we worked toward developing a suitable orthotopic animal model for nasal septum repair, while simultaneously evaluating the biocompatibility of marine collagen. Seeded and unseeded scaffolds were transplanted into nasal septum defects in an orthotopic rat model for 1, 4, and 12 weeks. Explanted scaffolds were histologically and immunohistochemically evaluated. Scaffolds did not induce any cytotoxic reactions in vitro. Chondrocytes were able to adhere to marine collagen and produce cartilaginous matrix proteins, such as collagen type II. Treating septal cartilage defects in vivo with seeded and unseeded scaffolds led to a significant reduction in the number of nasal septum perforations compared to no replacement. In summary, we demonstrated that marine collagen matrices provide excellent properties for cartilage tissue engineering. Marine collagen scaffolds are able to prevent septal perforations in an autologous, orthotopic rat model. This newly described experimental surgical procedure is a suitable way to evaluate new scaffold materials for their applicability in the context of nasal cartilage repair.

Processed xenogenic cartilage as innovative biomatrix for cartilage tissue engineering: effects on chondrocyte differentiation and function

One key point in the development of new bioimplant matrices for the reconstruction and replacement of cartilage defects is to provide an adequate microenvironment to ensure chondrocyte migration and de novo synthesis of cartilage-specific extracellular matrix (ECM). A recently developed decellularization and sterilization process maintains the three-dimensional (3D) collagen structure of native septal cartilage while increasing matrix porosity, which is considered to be crucial for cartilage tissue engineering. Human primary nasal septal chondrocytes were amplified in monolayer culture and 3D-cultured on processed porcine nasal septal cartilage scaffolds. The influence of chondrogenic growth factors on neosynthesis of ECM proteins was examined at the protein and gene expression levels. Seeding experiments demonstrated that processed xenogenic cartilage matrices provide excellent environmental properties for human nasal septal chondrocytes with respect to cell adhesion, migration into the matrix and neosynthesis of cartilage-specific ECM proteins, such as collagen type II and aggrecan. Matrix biomechanical stability indicated that the constructs retrieve full stability and function during 3D culture for up to 42 days, proportional to collagen type II and GAG production. Thus, processed xenogenic cartilage offers a suitable environment for human nasal chondrocytes and has promising potential for cartilage tissue engineering in the head and neck region.

Insulin-like Growth Factor-I and Growth Differentiation Factor-5 Promote the Formation of Tissue-Engineered Human Nasal Septal Cartilage

Tissue engineering of human nasal septal chondrocytes offers the potential to create large quantities of autologous material for use in reconstructive surgery of the head and neck. Culture with recombinant human growth factors may improve the biochemical and biomechanical properties of engineered tissue. The objectives of this study were to (1) perform a high-throughput screen to assess multiple combinations of growth factors and (2) perform more detailed testing of candidates identified in part I. In part I, human nasal septal chondrocytes from three donors were expanded in monolayer with pooled human serum (HS). Cells were then embedded in alginate beads for 2 weeks of culture in medium supplemented with 2% or 10% HS and 1 of 90 different growth factor combinations. Combinations of insulin-like growth factor-I (IGF-1), bone morphogenetic protein (BMP)-2, BMP-7, BMP-13, growth differentiation factor-5 (GDF-5), transforming growth factor β (TGFβ)-2, insulin, and dexamethasone were evaluated. Glycosaminoglycan (GAG) accumulation was measured. A combination of IGF-1 and GDF-5 was selected for further testing based on the results of part I. Chondrocytes from four donors underwent expansion followed by three-dimensional alginate culture for 2 weeks in medium supplemented with 2% or 10% HS with or without IGF-1 and GDF-5. Chondrocytes and their associated matrix were then recovered and cultured for 4 weeks in 12 mm transwells in medium supplemented with 2% or 10% HS with or without IGF-1 and GDF-5 (the same medium used for alginate culture). Biochemical and biomechanical properties of the neocartilage were measured. In part I, GAG accumulation was highest for growth factor combinations including both IGF-1 and GDF-5. In part II, the addition of IGF-1 and GDF-5 to 2% HS resulted in a 12-fold increase in construct thickness compared with 2% HS alone (p < 0.0001). GAG and type II collagen accumulation was significantly higher with IGF-1 and GDF-5. Confined compression modulus was greatest with 2% HS, IGF-1, and GDF-5. Supplementation of medium with IGF-1 and GDF-5 during creation of neocartilage constructs results in increased accumulation of GAG and type II collagen and improved biomechanical properties compared with constructs created without the growth factors.

Crushed Cartilage Grafts: Is Overcorrection Necessary?

Archives of Facial Plastic SurgeryVol. 10, No. 6 Free AccessCrushed Cartilage Grafts: Is Overcorrection Necessary?Ozcan CakmakOzcan CakmakCorrespondence: Dr Cakmak, Department of Otorhinolaryngology, Baskent University Hospital, 6. Cadde, Bahcelievler, 06490 Ankara, Turkey (E-mail Address: cakmak@baskent-ank.edu.tr)Search for more papers by this authorPublished Online:3 Nov 2008AboutSectionsPDF/EPUB Permissions & CitationsPermissionsDownload CitationsTrack CitationsAdd to favorites Back To Publication ShareShare onFacebookTwitterLinked InRedditEmail I read the article by Ale de Souza et alref-qle80002-1 with great interest. The authors concluded that crushed grafts had inferior results with lesser chondrocyte viability compared with noncrushed forms, and slight overcorrection was recommended during placement of crushed cartilage grafts. Based on our experimental and clinical findings, I believe the need of overcorrection is highly dependent on the level of crushing applied.Conflicting information exists regarding the viability of the crushed autogenous cartilage grafts and the predictability of their clinical outcomes. Recently, we attempted to resolve these inconsistent results by assessing the influence of the degree of crushing on the rabbit auricularref-qle80002-2 and human nasal septalref-qle80002-3 cartilages, and the clinical outcome of crushed grafts in rhinoplasty.ref-qle80002-4 Our experimental studiesref-qle80002-2,ref-qle80002-3 showed that although significant or severe crushing reduced chondrocyte viability and proliferation, slight or moderate crushing enabled good chondrocyte viability and proliferation rates comparable with those of intact cartilage. Our recent clinical seriesref-qle80002-4 confirmed findings from our previous studiesref-qle80002-2,ref-qle80002-3 that the degree of crushing applied is important for the resorption rates and long-term clinical outcome of crushed cartilage grafts used in rhinoplasty.Ale de Souza et alref-qle80002-1 crushed the rabbits' nasal septal cartilage with 2 moderate forced taps to “soften the surface without reducing the elastic strength of the cartilage”ref-qle80002-1(p251) and defined that grade of crushing as moderate. Although the criteria used by Ale de Souza et alref-qle80002-1 to define the level of crushing are very similar to those used by my group,ref-qle80002-2,ref-qle80002-3,ref-qle80002-4 their term “moderate” may lead to misinterpretations. The degree of crushing was previously defined in my group’s studies as follows: slightly crushed indicates 1 hit of moderate force to soften the surface without reducing cartilage elastic strength; moderately crushed, 2 moderate-force hits to soften the surface and also reduce elastic strength; significantly crushed, 3 to 4 moderate-force hits, enough to cause the graft to bend with gravity; and severely crushed, 5 to 6 forceful hits to totally destroy the integrity of the cartilage.On the one hand, based on our experimental results and clinical experience, significant or severe crushing reduces chondrocyte viability and proliferation and could result in an unpredictable degree of graft volume loss and should not be used as filler material. On the other hand, slight or moderate crushing of the autogenous cartilage produces an outstanding graft material that is effective in concealing irregularities, filling defects, and creating a smoother surface, with excellent long-term clinical outcome and predictable esthetic result without a need for overcorrection.Financial Disclosure: None reported. References Ale de Souza MM, Gregório LC, Sesso R, Souza SA, Settanni F. Study of rabbit septal cartilage grafts placed on the nasal dorsum. Arch Facial Plast Surg. 2008;10(4):250–254 18645093 Abstract, Google ScholarCakmak O, Bircan S, Buyuklu F, Tuncer I, Dal T, Ozluoglu LN. Viability of crushed and diced cartilage grafts: a study in rabbits.. Arch Facial Plast Surg. 2005;7(1):21–26 15655170 Abstract, Google ScholarCakmak O, Buyuklu F, Yilmaz Z, Sahin FI, Tarhan E, Ozluoglu LN. Viability of cultured human nasal septum chondrocytes after crushing.. Arch Facial Plast Surg. 2005;7(6):406–409 16301462 Link, Google ScholarCakmak O, Buyuklu F. Crushed cartilage grafts for concealing irregularities in rhinoplasty.. Arch Facial Plast Surg. 2007;9(5):352–357 17875829 Link, Google ScholarFiguresReferencesRelatedDetailsCited byA Classification for Degree of Crushed Cartilage Ozcan Cakmak and Hande Altintas1 November 2010 | Archives of Facial Plastic Surgery, Vol. 12, No. 6Rhinoplasty A View From the United Kingdom Julian M. Rowe-Jones2 November 2009 | Archives of Facial Plastic Surgery, Vol. 11, No. 6Rhinoplasty at the Global Crossroads Fazil Apaydin2 November 2009 | Archives of Facial Plastic Surgery, Vol. 11, No. 6 Volume 10Issue 6Nov 2008 InformationCopyright 2008 American Medical Association. All Rights Reserved. Applicable FARS/DFARS Restrictions Apply to Government Use.To cite this article:Ozcan Cakmak.Crushed Cartilage Grafts: Is Overcorrection Necessary?.Archives of Facial Plastic Surgery.Nov 2008.428-428.http://doi.org/10.1001/archfaci.10.6.4281Published in Volume: 10 Issue 6: November 3, 2008PDF download

The microscopic biological response of human chondrocytes to bovine bone scaffold

This study was performed to determine the microscopic biological response of human nasal septum chondrocytes and human knee articular chondrocytes placed on a demineralized bovine bone scaffold. Both chondrocytes were cultured and seeded onto the bovine bone scaffold with seeding density of 1 x 105 cells per 100 microl/scaffold and incubated for 1, 2, 5 and 7 days. Proliferation and viability of the cells were measured by mitochondrial dehydrogenase activity (MTT assay), adhesion study was analyzed by scanning electron microscopy and differentiation study was analyzed by immunofluorescence staining and confocal laser scanning electron microscopy. The results showed good proliferation and viability of both chondrocytes on the scaffolds from day 1 to day 7. Both chondrocytes increased in number with time and readily grew on the surface and into the open pores of the scaffold. Immunofluorescence staining demonstrated collagen type II on the scaffolds for both chondrocytes. The results showed good cells proliferation, attachment and maturity of the chondrocytes on the demineralized bovine bone scaffold. The bovine bone being easily resourced, relatively inexpensive and non toxic has good potential for use as a three dimensional construct in cartilage tissue engineering.

The Effects of Palmvitee on Human Nasal Septal Chondrocytes Culture Expansion and Cartilage Reconstruction

Palmvitee may act as a beneficial supplement to cartilage tissue engineering which is currently still has certain limitation on chondrocyte expansion and dedifferentiation. Our objectives in this study were to evaluate the effects of Palmvitee on the growth kinetic and phenotype gene expression of human nasal septal chondrocytes in monolayer expansion as well as cartilage reconstruction via tissue engineering technology. Human chondrocytes were cultured in mediums containing various Palmvitee concentrations. Among the different test groups, the medium containing Palmvitee at 3 µg/ml supported the highest chondrocyte growth rate. The gene expression in monolayer chondrocytes culture supplemented with 3 µg/ml Palmvitee demonstrated similar results as in control. Cultured chondrocytes from the medium added with 3 µg/ml Palmvitee were then mixed with Pluronic F127 for cartilage reconstruction in nude mice model. Palmvitee supplementation supported the engineered cartilage development as shown in the histological and gene expression analysis on engineered tissue after 8 weeks of implantation. Therefore, Palmvitee at 3 µg/ml is beneficial for the human nasal septal chondrocytes monolayer expansion and cartilage engineering.

Effect of bone morphogenetic proteins 2 and 7 on septal chondrocytes in alginate

To determine the effects of bone morphogenetic proteins (BMP)-2 and -7, and serum, on extracellular matrix production by human septal chondrocytes in alginate. Human nasal septal chondrocytes were expanded, suspended in alginate, and cultured in BMP-2 or 7, with and without serum. The optimal concentration of each growth factor was determined based on matrix production. Next, the synergistic effects of BMP-2 and -7 at optimal concentrations were determined on separate beads, based on matrix quantity and histology. Matrix content was highest with concentrations of BMP-2 and -7 of 100 ng/ml and 20 ng/ml, respectively, with serum. Adding both BMP-2 and -7, with serum, increased matrix content by factors of 5.1 versus serum-only cultures, 2.7 versus only BMP-2 with serum, and 2.4 versus only BMP-7 with serum. All comparisons were statistically significant. BMP-2 and -7 significantly increase production of extracellular matrix by septal chondrocytes suspended in alginate. The presence of serum improves matrix production. BMP-2 and -7 have great potential for use in cartilage tissue engineering.

The expansion potential of human nasal septum chondrocytes for the formation of engineered cartilage

The expansion potential of human nasal septum chondrocytes for the formation of engineered cartilage

Viability of Cultured Human Nasal Septum Chondrocytes After Crushing

Viability of Cultured Human Nasal Septum Chondrocytes After Crushing

Stored Human Septal Chondrocyte Viability Analyzed by Confocal Microscopy

To analyze the effects of prolonged storage time, at warm and cold temperatures, on the viability of human nasal septal chondrocytes and to understand the implications for tissue engineering of septal cartilage. Basic science. Septal cartilage was obtained from 10 patients and placed in bacteriostatic isotonic sodium chloride solution. Four specimens were kept at 23 degrees C, and 4 were kept at 4 degrees C. The viability of the chondrocytes within the cartilage was assessed using confocal laser scanning microscopy every 5 days. The 2 other specimens were assessed for viability on the day of harvest. Viability on the day of harvest was 96%, implying minimal cell death from surgical trauma. After 1 week, cell survival in all specimens was essentially unchanged from the day of harvest. At 23 degrees C, the majority (54%) of cells were alive after 20 days. At 4 degrees C, 70% of cells survived 1 month and 38% were alive at 2 months. Qualitatively, chondrocytes died in a topographically uniform distribution in warm specimens, whereas cold specimens displayed a more irregular pattern of cell death. Septal chondrocytes remain viable for prolonged periods when stored in simple bacteriostatic isotonic sodium chloride solution, and such survival is enhanced by cold storage.

Growth and Growth Factor Production by Human Nasal Septal Chondrocytes in Serum-Free Media

Tissue-engineered human cartilage offers vast possibilities as a source of graft implant material for reconstructive surgery. Serum-supplemented growth media is successful in supporting chondrocyte proliferation in vitro. Serum, however, contains exogenous growth factors that hamper the identification and quantification of growth factors autogenously produced by chondrocytes. We explore the possibility of using a commercially available serum-free medium UltraCULTURE as an alternative to modified Webber's medium (MWM), the standard media used in chondrocyte cell culture. Human nasal septal chondrocytes were grown in UltraCULTURE containing various concentrations of basic fibroblast growth factor (bFGF; 0, 1, 10, and 100 ng/mL) with or without insulin-like growth factor and compared with chondrocytes grown in MWM. Growth curves and transforming growth factor (TGF) beta 1 production were analyzed. We found no differences in the ability to sustain cell viability in culture between the two base media types. We also found no statistically significant differences in TGF-beta 1 production by chondrocytes grown in either system. Finally, there were no statistically significant differences in chondrocyte proliferation between cultures supplemented with bFGF at 10 and 100 ng/mL. UltraCULTURE media is a cost-effective, serum-free alternative to standard media with compatible growth characteristics. It offers specific advantages over standard serum-containing media for the precise measurement of autogenous growth factor production by cultured chondrocytes. Furthermore, UltraCULTURE's serum-free environment would be ideal for safely producing tissue-engineered cartilage grafts.