Biomedical subjects

Deborah Watson

Publications and source records attributed to Deborah Watson.

18 recordsLinked to original sources

PubMed2006

Stored human septal chondrocyte viability analyzed by confocal microscopy.

OBJECTIVES: 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. DESIGN: Basic science. SUBJECTS: 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. RESULTS: 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. CONCLUSION: Septal chondrocytes remain viable for prolonged periods when stored in simple bacteriostatic isotonic sodium chloride solution, and such survival is enhanced by cold storage.

Adult↗

PubMed2006

Human serum for tissue engineering of human nasal septal cartilage.

OBJECTIVE: To compare the chondrogenic and proliferative effects of pooled human serum (HS) and fetal bovine serum (FBS) on tissue-engineered human nasal septal chondrocytes. STUDY DESIGN AND SETTING: Human chondrocytes were expanded for one passage in monolayer in medium supplemented with 10% FBS, 2% HS, 10% HS, or 20% HS. Cells were then suspended in alginate beads for 3D culture for 2 weeks with 10% FBS, 2% HS, 10% HS, or 20% HS. RESULTS: Monolayer cell yields were greater with HS than FBS. In alginate, cellular proliferation, glycosaminoglycan production per cell, and type II collagen were significantly higher with 10% HS compared to 10% FBS controls. CONCLUSION: HS results in increased proliferation and production of cartilaginous extracellular matrix by tissue-engineered human nasal septal chondrocytes, compared to FBS controls. SIGNIFICANCE: Culture with human serum may facilitate creation of neocartilage constructs that more closely resemble native tissue.

Alginates↗

PubMed2005

Soft tissue reconstruction of the forehead and temple.

Soft tissue reconstruction of the forehead and temple challenges facial plastic surgeons to balance esthetic goals with functional concerns. Understanding the muscular and neurovascular anatomy is essential to achieve these ends. Reconstructive considerations include maintaining hairlines and eyebrows, minimizing scarring, using relaxed skin tension lines, and preserving motor and sensory function. Reconstructive options range from healing by secondary intention to primary closure, skin grafts, local flaps, island flaps, or any combination of these techniques.

Facial Injuries↗

PubMed2005

Submentoplasty.

Aging changes to the anterior neck can produce skin laxity, accumulation of submental fat, platysmal banding, and ptosis of the underlying neck structures. To rejuvenate the cervicomental region, the surgeon must accurately diagnose the anatomic irregularities and make appropriate recommendations for surgical correction. Because there are wide variations of anterior neck anatomy among individuals, it is necessary to "customize" the surgery in this area for each patient by employing single isolated procedures or multiple techniques. A stepwise approach in the management of the submentum can facilitate this goal and guide the surgeon in selecting the appropriate procedures for each individual.

Aging↗

PubMed2005

Characterization of human nasal septal chondrocytes cultured in alginate.

BACKGROUND: After serial passages in monolayer, chondrocytes dedifferentiate into a fibroblast-like phenotype. Our objective was to determine if culture in alginate affects the phenotype of dedifferentiated human nasal septal chondrocytes. STUDY DESIGN: Human nasal septal chondrocytes were seeded at low density and passaged in monolayer culture. At passages (P) 1, 2, and 3 a portion of cells were cultured in alginate. Collagen, glycosaminoglycan (GAG), and DNA production were assessed. RESULTS: Chondrocytes in alginate proliferated less yet produced higher levels of GAG and collagen than those in monolayer culture. Alginate encapsulated P1 chondrocytes stained strongly for GAG and collagen type II, and minimally for collagen type I. Monolayer cells at P0 and P1 stained positively for collagen type II. All monolayer passages stained positive for collagen type I with minimal GAG staining. CONCLUSIONS: Compared with monolayer culture, alginate stimulates deposition of GAG and collagen type II, and supports the chondrocyte phenotype through P1, but does not promote redifferentiation.

Alginates↗

PubMed2005

Growth and phenotype of low-density nasal septal chondrocyte monolayers.

OBJECTIVE: To analyze the growth patterns and differentiation of human septal chondrocyte monolayers of different seeding densities. STUDY DESIGN: Chondrocytes from 8 donors were plated at densities ranging from 20,000 cells/cm(2) (high density) to 300 cells/cm(2) (very low density). Confluency, cellularity, and glycosaminoglycan content were determined from days 1 to 15. RESULTS: Confluency was attained at 5.8, 8.3, 11.0, and 14.8 days for high-, intermediate-, low-, and very low-density monolayers, respectively (P < 0.001). Regression growth curves showed typical lag, logarithmic, and stationary phases. Confluent monolayers attained similar cellularity (power = 0.94) and differentiation (power = 0.88), regardless of initial density. CONCLUSIONS: Human septal chondrocyte monolayers reach confluency from very low initial densities. Growth patterns, cellularity, and differentiation are similar to other starting densities. SIGNIFICANCE: Very low-density monolayers expanded cell number 838-fold in 1 passage and therefore are sufficient for tissue-engineering purposes. This is important because of the requirement of maintaining differentiation and the limitation of small tissue harvest specimens.

Adult↗

PubMed2005

Effect of growth factors on cell proliferation, matrix deposition, and morphology of human nasal septal chondrocytes cultured in monolayer.

OBJECTIVES: Tissue engineering of septal cartilage provides ex vivo growth of cartilage from a patient's own septal chondrocytes for use in craniofacial reconstruction. To become clinically applicable, it is necessary to rapidly expand a limited population of donor chondrocytes and then stimulate the production of extracellular matrix on a biocompatible scaffold. The objective of this study was to determine favorable serum-free culture conditions for proliferation of human septal chondrocytes using various concentrations and combinations of four growth factors. STUDY DESIGN: Prospective, randomized, controlled study. METHODS: Nasal septal chondrocytes from six patient donors were isolated by enzymatic digestion and expanded in monolayer culture in both serum-free media (SFM) and 2% fetal bovine serum (FBS). Both of these groups were exposed to varying concentrations and combinations of transforming growth factor (TGF)-beta1, basic fibroblast growth factor (FGF)-2 both at 1, 5, and 25 ng/mL, and bone morphogenetic protein (BMP)-2 and insulin-like growth factor (IGF)-1, both at 5, 25, and 125 ng/mL in the medium during the expansion phase. Cell morphology was assessed throughout the culture duration. After 7 days of monolayer growth, cultures were assessed for cellularity and glycosaminoglycan (GAG) content. RESULTS: The addition of low-dose FBS in culture media consistently led to significantly greater cell proliferation and matrix deposition than the SFM cell cultures. FGF-2 and TGF-beta1 both alone and in combination led to the greatest proliferative effect compared with the other growth factors. In contrast, BMP-2 and IGF-1 led to the least cell proliferation although was most effective in retaining chondrocyte cell morphology. CONCLUSIONS: With the addition of TGF-beta1 and FGF-2 to culture media, the concentration of serum can be greatly decreased and possibly eliminated altogether without jeopardizing cell proliferation.

Adolescent↗

PubMed2004

Review of injectable materials for soft tissue augmentation.

Although a variety of injectable agents are available for soft tissue augmentation, no perfect filler material exists. The advantages and disadvantages of commonly used materials such as bovine collagen have been well documented. However, many newer injectables are still undergoing experimentation to determine their clinical efficacy and long-term safety. This article details recent scientific work to compare injectable materials from the following categories: xenografts (bovine collagen and hyaluronic acid derivatives), autografts (autologous fat, Isolagen, and Autologen), homografts (Dermalogen and Cymetra), and synthetic materials (fluid silicone and Artecoll).

Adipose Tissue↗

PubMed2004

Repair of the torn earlobe.

There exists a variety of techniques to repair torn earlobes. Some of these methods preserve the original earring hole during the earlobe repair, but many authors advocate complete closure of the torn earlobe and repiercing it at a later time. This article reviews the published techniques for earlobe repair. Illustrations accompany some of the different methods, and helpful recommendations are listed for a surgical approach to torn earlobes.

Cosmetic Techniques↗

PubMed2004

Microdermabrasion: a review.

Microdermabrasion is used by clinicians and aestheticians worldwide to rejuvenate facial skin. The microdermabrator contains aluminum oxide or sodium chloride crystals that strike the skin and produce superficial trauma. It is theorized that the repetitive intraepidermal injury causes gradual improvement in damaged skin by stimulating fibroblast proliferation and collagen production, leading to new collagen deposition in the dermis. In this review of microdermabrasion, we discuss studies performed to elucidate the mechanism of action by which microdermabrasion promotes facial resurfacing. We also discuss other existing techniques for facial rejuvenation and resurfacing.

Dermabrasion↗

PubMed2004

Tissue-engineered human nasal septal cartilage using the alginate-recovered-chondrocyte method.

OBJECTIVES: Tissue engineering of nasal septal cartilage has numerous potential applications in craniofacial reconstruction. Chondrocytes suspended in alginate gel have been shown to produce a substantial cell-associated matrix. The objective of this study was to determine whether cartilage tissue could be generated using the alginate-recovered-chondrocyte (ARC) method, in which chondrocytes are cultured in alginate as an intermediate step in tissue fabrication. METHODS: Nasal septal chondrocytes from five patient donors were isolated by enzymatic digestion, then expanded in monolayer culture. At confluency, a portion of those cells were seeded at high density onto a semipermeable membrane and cultured for 14, 21, or 28 days (monolayer group). The remaining cells were suspended in alginate and cultured until a cell-associated matrix was observed (10-17 days). Cells and their associated matrix were released from alginate (ARC group), seeded onto a semipermeable membrane, and cultured as already described. DNA (Hoechst 33258 Assay), glycosaminoglycan (GAG; dimethylmethylene blue assay), and collagen (hydroxyproline assay) were analyzed biochemically. Immunohistochemistry was performed to assess expression of collagens type I and type II. Histochemistry was performed to localize cells accumulating sulfated GAG (Alcian blue stain). RESULTS: The ARC constructs, in contrast to the monolayer constructs, had substantial structural stability and the histologic and gross appearance of cartilaginous tissue. ARC constructs demonstrated significantly greater GAG and collagen accumulation than monolayer constructs (P <.05). Histologic analysis revealed substantial GAG and collagen type II production and only moderate collagen type I production. The composition of the matrix was thus similar to that of native human septal cartilage. CONCLUSIONS: Tissue-engineered human nasal septal cartilage using the ARC method has the histologic and gross appearance of native cartilage and has biochemical composition more like that of native cartilage than monolayer constructs. This is the first report of human nasal septal neocartilage formation without the use of biodegradable scaffolds.

Alginates↗

PubMed2003

Polymorphisms within the vitamin B12 dependent methylmalonyl-coA mutase are not risk factors for neural tube defects.

Methionine synthase and methylmalonyl-CoA mutase (mutase) are the only two known vitamin B(12) (B(12)) dependent enzymes in humans. A lower level of B(12) has been shown to be an independent maternal risk factor for neural tube defects (NTDs) prompting an investigation of common genetic variants within B(12) dependent enzymes. To investigate the role of methylmalonyl-CoA mutase variants we studied 279 complete NTD triads (NTD affected case and both parents) and 256 controls. Based on case-control and family based (transmission disequilibrium test) analyses we did not find an association between the mutase single nucleotide polymorphisms (SNPs) K212K (636A-->G), H532R (1595A-->G) and V671I (2011G-->A) and NTDs. However, there was a significant difference in the frequencies of these polymorphisms between a group of African Americans and American Caucasians (K212K, P=0.002; H532R, P</=0.001; V671I, P=0.006). In conclusion, common variants in the mutase gene do not appear to be risk factors for NTDs but their allele frequencies are significantly different between ethnic groups.

Case-Control Studies↗

PubMed2003

Human septal chondrocyte redifferentiation in alginate, polyglycolic acid scaffold, and monolayer culture.

OBJECTIVES/HYPOTHESIS: Tissue engineering laboratories are attempting to create neocartilage that could serve as an implant material for structural support during reconstructive surgery. One approach to forming such tissue is to proliferate chondrocytes in monolayer culture and then seed the expanded cell population onto biodegradable scaffolds. However, chondrocytes are known to dedifferentiate after this type of monolayer growth and, as a result, decrease their production of cartilaginous extracellular matrix components such as sulfated glycosaminoglycans. The resultant tissue lacks the biomechanical properties characteristic of cartilage. The objective of the study was to determine whether different culture systems could induce monolayer-expanded human septal chondrocytes to redifferentiate and form extracellular matrix. STUDY DESIGN: Laboratory research. METHODS: Chondrocytes were isolated from human nasal septal cartilage of five donor patients (age, 35.8 +/- 9.3 y). Cell populations were seeded at low density (30,000 cells/cm2) into monolayer culture and expanded for 4 to 6 days. Following trypsin release, chondrocytes were placed into three different systems for neocartilage formation: alginate beads, polyglycolic acid scaffolds, and monolayer. After 7 and 14 days of growth, neocartilage was analyzed using histological and quantitative biochemical assessment of cellularity (Hoechst 33258 assay) and sulfated glycosaminoglycan content (dimethyl methylene blue assay). RESULTS: Histologically, alginate beads contained spherical chondrocytes surrounded by dense extracellular matrix, an appearance similar to that of native cartilage. In contrast, polyglycolic acid scaffolds and monolayer cultures contained elongated cells with scant staining for matrix sulfated glycosaminoglycans, which are features that are characteristic of dedifferentiated chondrocytes. Biochemical analysis demonstrated a lower level of cell proliferation (P <.001) in scaffolds (+52% over baseline) and alginate (+96% over baseline) than in monolayer (+366% over baseline), as well as a higher content of sulfated glycosaminoglycans per cell (P <.001), after 14 days of growth in alginate culture than in either polyglycolic acid scaffolds (19-fold difference) or monolayer (98-fold difference). CONCLUSIONS: Of the systems compared, monolayer-expanded human septal chondrocytes demonstrated the greatest accumulation of sulfated glycosaminoglycans per cell when grown in alginate beads. Future research on cartilage tissue engineering may use alginate culture for reverting dedifferentiated cells back to the chondrocytic phenotype.

Alginates↗

PubMed2002

Torn earlobe repair.

A variety of techniques have been described in the literature to repair torn earlobes. Some of these methods incorporate the reconstruction of the earring hole during earlobe repair, but many authors still recommend repiercing the earlobe at a later time. The present article is a comprehensive review of the published techniques for earlobe repair. Illustrations are provided to facilitate the description of the different methods and helpful recommendations are listed for a surgical approach to torn earlobes.

Ear↗

PubMed2002

Effects of serial expansion of septal chondrocytes on tissue-engineered neocartilage composition.

OBJECTIVES: Cartilage grafts for reconstructive surgery may someday be created from harvested autologous chondrocytes that are expanded and seeded onto biodegradable scaffolds in vitro. This study sought to quantify the biochemical composition of neocartilage engineered from human septal chondrocytes and to examine the effects of cell multiplication in monolayer culture on the ultimate composition of the neocartilage. METHODS: Human septal chondrocytes from 10 donors were either seeded immediately after harvest (passage 0 [P(0)]) onto polyglycolic acid (PGA) scaffolds or underwent multiplication in monolayer culture before scaffold seeding at passage 1 (P(1)) and passage 2 (P(2)). Cell/scaffold constructs were grown in vitro for 7, 14, and 28 days. Neocartilage constructs underwent histologic analysis for matrix sulfated glycosaminoglycan (S-GAG) and type II collagen as well as quantitative assessment of cellularity (Hoescht 33258 assay), S-GAG content (dimethylmethylene blue assay), and collagen content (hydroxyproline assay). RESULTS: Histologic sections of constructs seeded with P(0) cells stained strongly for S-GAG and type II collagen, whereas decreased staining for both matrix components was observed in constructs derived from P(1) and P(2) cells. Cellularity, S-GAG content, and total collagen content of constructs increased significantly from day 7 to day 28. S-GAG accumulation in P(0) constructs was higher than in either P(1) (P < 0.05) or P(2) (P < 0.01) constructs, whereas cellularity and total collagen content showed no difference between passages. CONCLUSION: Neocartilage created from chondrocytes that have undergone serial passages in monolayer culture exhibited decreased matrix S-GAG and type II collagen, indicative of cellular dedifferentiation. SIGNIFICANCE: The alterations of matrix composition produced by dedifferentiated chondrocytes may limit the mechanical stability of neocartilage constructs.

Adult↗

PubMedDate not supplied

A compositional analysis of human nasal septal cartilage.

BACKGROUND: Nasal septal cartilage is well established as an autograft material. Tissue engineering methods are now being developed to synthesize cartilage constructs with the properties of this type of cartilage. However, important baseline data on the composition of native septal cartilage is sparse. OBJECTIVES: To characterize quantitatively the major biochemical constituents of native adult human septal cartilage and determine age- or sex-related variation in composition. METHODS: Cartilage was harvested from the inferior region of the nasal septum in 33 patients (mean +/- SD age, 47.0 +/- 13.5 years; range, 24-80 years) during routine septoplasty or septorhinoplasty. Biochemical assays were used to determine the quantities, relative to wet weight, of the major constituents of cartilage: water, collagen (from hydroxyproline), sulfated glycosaminoglycan (sGAG), and chondrocytes (from DNA). RESULTS: On average, each gram of wet cartilage contained 77.7% water, 7.7% collagen, 2.9% sGAG, and 24.9 million cells. Hydration and collagen content showed no significant age variation. Advancing age was associated with a reduction in sGAG content (7.7% per decade, P =.02) and cellularity (7.4% per decade, P =.05). No significant sex differences were found in any of these cartilage constituents. CONCLUSIONS: This study represents the first biochemical characterization of the composition of native human septal cartilage. The data serve as a baseline for future comparison of the properties of tissue-engineered neocartilage constructs. Furthermore, the age-associated variations in cartilage composition have implications for patient selection for reconstructive procedures.

Adult↗

PubMedDate not supplied

Compressive biomechanical properties of human nasal septal cartilage.

BACKGROUND: Nasal septal cartilage is frequently used in nasal reconstruction and is a common source of chondrocytes for cartilage tissue engineering. The biomechanical properties of septal cartilage have yet to be fully defined and this limits the ability to compare it to the various alternative tissue-implant materials or tissue-engineered neocartilage. Given the unique structure and orientation of the septum within the nose, we sought to investigate anisotropic behaviors of septal cartilage in compression and correlate this to the concentration of glycosaminoglycans (GAG) and collagen within the cartilage. METHODS: Human nasal septal cartilage specimens were tested in confined compression, with each sample analyzed in a medial orientation and also either a vertical or caudal-cephalic orientation, with the order of tests randomized. The equilibrium confined compression (aggregate) modulus, HAO, and the permeability, kp, at different offset compression levels were obtained for each compression test. After testing, the cartilage samples were solubilized, and the concentrations of GAG and collagen were obtained. RESULTS: Forty-nine compression tests (24 medial, 12 vertical, 13 caudal-cephalic) were run on cartilage specimens obtained from 21 patients. There was a significant effect of orientation on compression modulus, HAO, with the vertical (0.7 +/- 0.12 MPa) and caudal-cephalic (0.66 +/- 0.01 MPa) orientations being significantly stiffer (p = 0.05) than the medial orientation (0.44 +/- 0.04 MPa). There was a trend of an orientation effect on kp at 15% offset compression (p = 0.12) and a borderline significant effect of orientation on kp at 30% offset compression (p = 0.05), demonstrating the M orientation to be more permeable than both the vertical and caudal-cephalic orientations. Both univariate and multivariate analysis did not demonstrate a significant effect of order of compression, age, gender, thickness, dry/wet weight, GAG, or collagen on either HAO, or kp values (p > 0.05). CONCLUSION: This study provides new information on the compressive properties of septal cartilage along different axes of compression. The results demonstrate that human septal cartilage is anisotropic; the compressive stiffness is higher in the vertical and caudal-cephalic orientations than in the medial orientation. Additionally, the medial orientation tends to have the greatest permeability. The data obtained in this study provide a reference to which various craniofacial reconstruction materials and tissue-engineered neocartilage can be compared.

Adolescent↗

PubMedDate not supplied

Tensile biomechanical properties of human nasal septal cartilage.

BACKGROUND: The biomechanical properties of human septal cartilage have yet to be fully defined and thereby limits our ability to compare tissue-engineered constructs to native tissue. In this study, we analyzed the tensile properties of human nasal septal cartilage with respect to axis of tension, age group, and gender. METHODS: Fifty-five tensile tests were run on human septal specimens obtained from 28 patients. Samples obtained in the vertical and anterior-posterior (both above and within the maxillary crest) axes were subjected to equilibrium and dynamic tensile testing. RESULTS: The average values for strength, failure strain, equilibrium modulus and dynamic modulus were not found to be significantly different with respect to axis of tension testing, age group, or gender. Tensile results for septal cartilage were as follows: equilibrium modulus 3.01 +/- 0.39 MPa, dynamic modulus 4.99 +/- 0.49 MPa, strength 1.90 +/- 0.24 MPa, and failure strain 0.35 +/- 0.03 mm/mm. CONCLUSION: We confirm that septal cartilage has weaker tensile properties compared to articular cartilage and found no difference in strength with respect to age, gender, or axis of tension (isotropic).

Adult↗