Pedagogy on Gingival Fibroblast

CONTENTS
Introduction Embryology Structure Functions Heterogeneity of fibroblasts Implication of gingival fibroblast subtypes in health, disease and treatment Conclusion References

Introduction
Fibroblast Fibroblast is a type of cell that synthesizes the extra cellular matrix and structural components and plays an important role in wound healing Hence these cells are called Architects and care takers of connective tissues

Embryology
Fibroblasts are believed to arise from three distinct cellular origins: (Postlethwaite et al., 2004) PRIMARY MESENCHYME o It is widely accepted that the principle origin for fibroblast like all the other connective tissue cells is from primary mesenchymal cells and that upon appropriate stimulation, fibroblasts can proliferate to generate new fibroblasts o It demonstrates Vimentin Intermediate filament-associated proteins which are usually present in mesenchymal cells LOCAL EPITHELIALMESENCHYMAL TRANSITION (EMT) o Local EMT is a central mechanism for diversifying cells in the formation of complex tissues (Kalluri and Neilson, 2003). Fibroblasts can be derived by this

process in adult tissue following epithelial stress such as inflammation or tissue injury o A combination of cytokines associated with digestion of the basement membrane is believed to instigate EMT as well as matrix metalloproteinases (MMPs) with transforming growth factor beta (TGF-b) thought to be important in the induction of EMT (Zeisberg et al., 2001, 2003). BONE MARROW-DERIVED PRECURSORS o More recently, it has been suggested that fibroblasts can be derived from bone marrow-derived, precursor cells, which circulate in the blood termed fibrocytes (Phillips et al., 2004). These cells represent B0.05% of circulating blood cells that are believed to arise from CD14 peripheral blood monocytes. o All of these processes appear to contribute to the pathological accumulation of fibroblasts. Prior to tooth eruption, the gingival ridge and later the gum pads mark the sites of tooth crown penetration through the mucosa. The fibroblasts located in the gum pads and in the fully developed lamina propria of the gingiva probably arise from cells in the perifollicular mesenchyme.

Identification and Structure of Fibroblast

Fibroblasts are traditionally identified by their spindle shaped morphology and their ability to adhere to plastic in vitro (Tarin and Croft, 1969) They appear rounded, With a spherical nucleus in the centre and typical prolongations

They have variable activity based on which they are staged into Fibroblast : o involved in active secretory process o elliptical raised speckled nucleus with one or two nucleoli o abundant RER Fibrocyte : o less active and concerned with maintenance and tissue metabolism o small spindle shaped o relative less amount of RER

Functions
Fibroblasts are dynamic cells which performs specific functions within and between tissue types. production of structural elements o collagen o reticular fibres o elastic fibres production of extra cellular matrix o glycosaminoglycans o glycoproteins o Fibronectin etc. tissue turnover by o matrixmetalloproteinases

o phagocytosis Wound healing: Fibroblastic cells present in granulation tissue are known to exhibit morphologic and biologic properties intermediate between those of fibroblasts and those of smooth-muscle cells (Hakkinen and Larjava, 1992). The name myofibroblast was proposed for these cells, suggesting that they provide the force needed for wound contraction (Darby et al., 1990; Gabbiani, 1992, 1993).Interferon- has an antifibrotic effect, which probably results from its ability to inhibit fibroblast differentiation into myofibroblasts host defence o It has now been suggested that fibroblasts not only play an active role in defining the structure of tissue microenvironments but also modulate immune-cell behaviour by conditioning the local cellular and cytokine production. Therefore, the response of the microenvironment can be specifically tailored to the cause of the damage. o Fibroblast activation leads to production of cytokines TGF-, interleukin (IL)-8 and IL-10, chemokines and prostanoids such as prostaglandin E2 (Smith et al., 1997). o Fibroblasts can also regulate the behaviour of haematopoietic cells present in damaged tissue via CD40CD40 ligand interaction to synthesize high levels of IL-6, IL-8, cyclooxygenase-2 and the polysaccharide hyaluronan (Zhang et al., 1998; Pap et al., 2000) o Recent data demonstrate that fibroblasts display activation and interaction proteins (CD40, class II MHC, integrins) which permit cell-cell interaction with immune cells and subsequent cytokine production

Heterogeneity Of Fibroblasts
Studies of fibroblast heterogeneity date back to the early 1960s. Castor et al. (2) examined differences in fibroblasts cultured from dermis, articular tissues, mesothelial surfaces and periosteum. They found that mesothelial surface fibroblasts had a slower proliferative rate than fibroblasts from other tissues and produced more acid mucopolysaccharide than other fibroblast lines More evidence for fibroblast heterogeneity from cells derived from different anatomical sites has been shown in comparisons of orbital and dermal fibroblasts. Orbital fibroblasts play a role in the development of the ophthalmopathy associated with Grave's disease Based on their gene and protein expression (Parsonage et al., 2003). These have been named o Typical (peripheral) and o Atypical (lymphoid) fibroblasts. Studies of fibroblast heterogeneity date back to the early 1960s. Castor et al. (2) examined differences in fibroblasts cultured from dermis, articular tissues, mesothelial surfaces and periosteum. They found that mesothelial surface fibroblasts had a slower proliferative rate than fibroblasts from other tissues and produced more acid mucopolysaccharide than other fibroblast lines More evidence for fibroblast heterogeneity from cells derived from different anatomical sites has been shown in comparisons of orbital and dermal fibroblasts. Orbital fibroblasts play a role in the development of the ophthalmopathy associated with Grave's disease

Studies by Mollenhauer & Bayreuther observed 3 distinct fibroblast cell types based on morphology and proliferative characteristics, which displayed heterogeneity with respect to collagen production (type I and type III). o Fibroblasts termed FI produced very little type I and type III collagen. FII cells synthesized slightly more collagen and FIII cells produced large quantities of collagen

In spite of their similar appearance, fibroblasts are in fact a highly diverse population of cells exhibiting a considerable degree of both inter- and intra-site heterogeneity (as reviewed by Schor and Schor, 1987).

Differences between PDL and gingival fibroblast PDL fibroblast Gingival Fibroblast

Glycogen pools with in the cytoplasm with -----many bands of contractile microfilaments (actin three-fold higher in PDLF) Expresses smooth-muscle actin and myosin, Expresses only smooth-muscle actin Produces more amounts of GAGs and type I Releases larger amounts of PGE2 in response and type III collagen to bradykinin and histamine Produces more amount alkaline phosphatase ------suggestive of 'osteoblast-like' properties. High proliferation rate than PDLG PDLF possess the ability for mineralization (Ohshima et al., 1988) and for producing mineral-like nodules in vitro (Arceo et al., 1991). Gelatin, laminin, and vitronectin enhanced the attachment of PDG F. -------

collagen types I and IV promoted the attachment of GF,

Irwin and colleagues dissected the lamina propria of attached human gingiva into the papillary tips and deeper, reticular layers and demonstrated that fibroblasts from each layer differ in several ways

Papillary fibroblast
proliferated faster in primary culture small and spindly were similar to fetal skin fibroblasts in their production of a migration stimulating factor (MSF)

Reticular fibroblast
Relatively slow proliferation larger and more spread morphology similar to adult skin fibroblasts that do not produce MSF

The fetal-like gingival papillary fibroblasts may thus contribute to migration and beneficial, non-scarring wound healing in the oral mucosa. In the embryos, there is no conversion of fibroblasts to myofibroblasts and the angiogenic response is also of a lesser magnitude. Compared with wounds in adult tissues, wounds in embryos show low but transient levels of expression of TGF 1 and since TGF 1 has been implicated in pathofibrotic conditions, this can explain the tendency for adult wounds to show scarring and contraction. Antibodies that neutralize the effects of TGF 1 in healing wounds have been shown to reduce scarring when delivered to wounds

Bordin & Page demonstrated heterogeneity in the expression of Clq receptors on gingival fibroblasts. The fact that these subsets could be repeatedly identified, fractionated and cultured with no loss of phenotype most strongly supported the concept of gingival fibroblast heterogeneity

fibroblast heterogeneity in expression of other surface markers such as CD4,CD40 and CD60

Implications Of Gingival Fibroblast Subtypes In Health Disease And Treatment

Identification of these cells in health and disease will allow a determination of their significance in wound repair. Fibroblasts from gingiva derived from periodontal disease patients show an increase in mRNA for type I collagen trimer. Nicotine alters fibroblast attachment and integrin expression(beta 1) and decrease collagen production while increasing collagenase production Differences in the response to cyclosporine A have been reported to be heterogeneous among gingival fibroblasts about 35% of the gingival fibroblasts failed to bind cyclosporin A, with only the binding subset proliferating in response to the drug. The CD4 fibroblast subpopulation in gingival and PDL tissues may be infected with HIV and be deleted in a manner similar to CD4 T lymphocytes during AIDS. Loss or perturbation of this fibroblast subset could be implicated in the lesions manifested in the oral cavity in HlV-associated periodontal disease. The loss of fibroblasts in severely inflamed periodontal tissue may be a consequence of a fibroblast subset(s) being deleted from these tissues. The resulting loss of protein and extracellular matrix production by these cells would lead to diminished tissue integrity and loss of tooth support. One emergent concept is that a small pool of progenitor fibroblasts exists in tissue, which is responsible for tissue turnover and repair. Further studies of the functional significance of specific subsets will pinpoint their role in the pathologic changes which occur in periodontal disease

Such therapies may include Stimulating growth and differentiation receptors on fibroblasts or Seeding damaged tissue with fibroblast progenitor cells. Exploring how the fibroblast interacts with host defences will undoubtedly reveal new information about the pathogenesis of periodontal disease Pathological alterations of oral connective tissues may result from a clonal imbalance of resident fibroblast subtypes rather than from the presence of abnormal or cytopathologically altered cells. Furthermore, the relative proportions of various subtypes and their functional activities in the tissue can be regulated by specific cellular interactions with molecules such as cytokines, many of which are present in the systemic circulation (Saito et al., 1990).

Conclusion
Thus I conclude my pedagogy by saying that study of gingival fibroblast heterogeneity will make great contributions to the understanding of pathologic processes and wound healing. When the role of various fibroblast subsets is understood manipulation of the response of these cells to injury and environmental challenge may be possible, leading to new and specific therapies for periodontal disease and other connective tissue diseases.

References
1. REVIEW Fibroblasts as novel therapeutic targets in chronic inflammation SJ Flavell, TZ Hou, S Lax, AD Filer, M Salmon and CD Buckley British Journal of Pharmacology (2008) 153, S241S246 2. Functional Characteristics of Gingival and Periodontal Ligament Fibroblasts C. Giannopoulou and G. Cimasoni J Dent Res 75(3): 895-902, March, 1996 3. Role of fibroblast subpopulations in periodontal physiology and pathology McCulloch CAG, Bordin S J Periodont Res 1991; 26: 144-154. 4. Fibroblast heterogeneity in the periodontium and other tissues Phipps RP, Borrello MA, Blieden TM J Periodont Res 1997; 32: 159-165.

5. Functional Characteristics of Gingival and Periodontal Ligament Fibroblasts C. Giannopoulou and G. Cimasoni J Dent Res 75(3): 895-902, March, 1996 6. A Comparative Study of Human Periodontal Ligament Cells and Gingival Fibroblasts in vitro M.J. SOMERMAN, S.Y. ARCHER', G.R. IMM2, and R.A. FOSTER J Dent Res 67(1):66-70, January, 1988 7. Is Fibroblast Heterogeneity Relevant To the Health, Diseases, and Treatments of Periodontal Tissues? P.C. Lekic, N. Pender and C.A.G. McCulloch CROBM 1997 8: 253