Faculty Bibliography

Fibroblast cultures derived from uninjured and reparative rabbit buccal mucosa were compared in terms of extracellular glycosaminoglycan (GAG) content and cellular response to interleukin-1 (IL-1). Under identical growth conditions, proliferation of both cell lines was the same. Both lines incorporated [3H]-glucosamine into GAG in cellular, pericellular, and medium fractions, with the majority of incorporated label residing in the medium. Dermatan sulfate (DS) was the predominant GAG in the medium fraction of both normal and wound fibroblast cultures; however, the two cell lines differed in the identity of the medium fraction's secondary GAG: chondroitin sulfate (CS) for normal fibroblasts and hyaluronic acid (HA) for wound-derived cells. The GAG content of the pericellular matrix for all cultures was the same regardless of the tissue of origin: heparan sulfate (HS) accompanied by a very small amount of CS. Exposure to IL-1 produced limited but highly specific effects: It was not mitogenic for either cell line but did cause a quantitative change (increase) in overall incorporation into GAG for medium and pericellular fractions for both cell lines. Further, IL-1 induced a qualitative change in GAG composition for normal mucosal fibroblastic medium fractions by causing the synthesis/release of heparan sulfate (HS) and a variant form of DS. These data support the hypothesis that different fibroblastic substrains can populate a given oral site as a function of variables such as injury and/or healing status

The glycosaminoglycan (GAG) content of rabbit skin, oral mucosa, and cultured [3H]-glucosamine-labeled dermal and submucosal fibroblasts was compared. Skin contained predominantly dermatan sulfate (DS) and a small amount of hyaluronic acid (HA), whereas mucosa contained primarily keratan sulfate (KS) and smaller quantities of HA and DS. Culture medium from dermal and submucosal fibroblasts contained GAGs co-electrophoresing with DS, HA, and chondroitin sulfate (CS), although the relative proportions of these GAG differed. CS isolated from dermal and mucosal fibroblast culture medium co-electrophoresed with chondroitin 4-sulfate (C4-S) on cellulose acetate, whereas dermal medium CS was resistant to digestion by chondroitinase ABC, and mucosal medium CS was chondroitinase ABC-susceptible. The pericellular matrix of dermal fibroblasts contained primarily DS and C4-S/C6-S, as confirmed by chondroitinase ABC digestion; the corresponding fraction of mucosal fibroblasts contained HS and a GAG co-electrophoresing with a C6-S standard, yet resistant to digestion by chondroitinase ABC. Thus the GAG content of dermal and mucosal fibroblasts differed both qualitatively in terms of the type of GAG secreted into the culture medium and pericellular matrix, and quantitatively, in terms of the relative proportions of these GAGs in both fractions. These differences support the concept of distinctive fibroblastic subpopulations in skin and mucosal tissue, inasmuch as the cells were subjected to identical culturing conditions

This report describes the healing of cutaneous wounds in experimental animals grafted with collagen-glycosaminoglycan (GAG) matrix/Silastic (Dow Corning Corp., Midland, MI) bilayers; assesses the feasibility of using collagen-GAG matrix as a vehicle for delivering culture-selected, autogenous fibroblasts to cutaneous wound sites; and evaluates the use of collagen-GAG/Silastic bilayers as mucosal substitutes. Cutaneous and mucosal wounds in New Zealand white rabbits were grafted with either acellular collagen-GAG/Silastic membrane or collagen-GAG/Silastic membrane previously seeded with cultured autogenous fibroblasts. Over 63 days, wound sites were analyzed at intervals based on wound contraction and histology. Cutaneous wound successfully incorporated grafted collagen-GAG matrix and were significantly inhibited in their rate and extent of wound contraction. Seeding membrane matrices with autogenous, cultured fibroblasts before grafting caused a marked increase in cellularity that persisted throughout the postgraft period. In mucosa, matrices were exteriorized rather than incorporated. This work suggests that collagen-GAG/Silastic bilayer may have value as a dermal substitute and, more significantly, may be appropriate as a vehicle for delivering culture-selected fibroblasts to cutaneous wound sites

Fibroblast cultures established from explants of mature scar and skin tissue were analyzed with regard to extracellular glycosaminoglycan (GAG) composition and response to interleukin-1 (IL-1). Following a serum-free 48 hour label with [3H]glucosamine, pericellular and medium GAGs were isolated by precipitation with cetylpyridinium chloride (CPC) and analyzed by cellulose acetate electrophoresis. In addition, susceptibility of the precipitates to Streptomyces hyaluronidase, chondroitinase ABC and heparitinase was determined. Labeled conditioned medium from the scar-derived cells contained both dermatan sulfate (DS) and hyaluronate (HA), as compared to medium from the control (skin-derived) cells which contained predominantly DS. IL-1 induced the appearance of chondroitin 4-sulfate (C4-S) in the medium of the scar cells with no concurrent effect on either DS or HA, and increased the amount of HA in the medium fraction of normal skin cells. The pericellular fraction of the scar-derived cells contained chondroitin 6-sulfate (C6-S) and DS; addition of IL-1 resulted in a shift from DS to heparan sulfate (HS), and the emergence of a pericellular GAG profile similar to that of normal dermal fibroblasts

Both clinical and empirical evidence suggests that the 'normal' temporomandibular joint produces noise during function. The purpose of this study was to determine the conditions under which these noises might arise. Joint sounds and mandibular movements were recorded simultaneously from 200 adults who had no previous history or present symptoms of TMJ pain or dysfunction. The joint sounds were recorded bilaterally by means of two separate miniature vibration transducers mounted on a common headband, and incisal point mandibular movements were measured by a magnetometer tracking system. Recordings were made while each subject opened and closed the mouth, first in a natural and comfortable manner, and then to maximum displacement. Two types of measurements were made: the appearance, onset, and duration of joint-propagated sounds in relation to relative mandibular position; and the spectral properties of the detected sounds. The results of this study showed that for natural opening and closing movements of the mandible, the 'normal' TMJ was silent for all age groups. However, for the maximum displacement condition, detectable sounds appeared at the points of maximum displacement in both the opening and closing phases of the cycle in over 80 percent of the subjects. However, these sounds were, as a class, substantially longer than, and spectrally distinct from, abnormal joint sounds, suggesting that they arise from a distinct physical substrate