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Original Article Protective effects of mesenchymal stem cell cond tional medium against inflammatory injury on human gingival fibroblast

Original Article Protective effects of mesenchymal stem cell cond tional medium against inflammatory injury on human gingival fibroblast

HGFs play crucial roles in the formation and regeneration of gingival interface of tooth im- plants. At the early stage of gingival interface healing, HGFs secrete thread-like compounds to cover the surface of tooth implant to facili- tate trauma repair via adhesion. After healing of tooth implants, HGFs maintain normal func- tion of biological closed interface, and parti- cipate in repair and reconstruction of tooth implants to achieve closure of tooth [13, 14]. Therefore, effective adhesion of HGFs and pro- liferation play critical roles in repairmen of peripheral inflammation around tooth implant. Currently abundant studies have been per- formed regarding IL-6 and periodontitis. Per- iodontal tissue can produce IL-6, which further destructs periodontal tissues via facilitating T lymphocyte differentiation and inhibit fibroblast growth, thus inducing pathological bone reab- sorption in periodontitis and playing important roles in inflammatory injury of periodontal tis- sues [15, 16]. MSCCM has different effects on fibroblast with different origins, as it has minor effects on normal fibroblast, but significantly suppress proliferation of scar-forming fibrobl- ast via changing extracellular matrix constru- cts by TGF-β/Smad signal pathway, and fight against oxidative stress injury of H9C2 cells by activating MAPK, PI3K/Akt signal pathway

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Original Article Effects of cigarette smoke condensate and nicotine on growth status and cytokine expression of human gingival fibroblast on titanium plate

Original Article Effects of cigarette smoke condensate and nicotine on growth status and cytokine expression of human gingival fibroblast on titanium plate

Smoking has become a major health concern for people’s health. Toxic substances such as nicotine, acrolein, and cotinine can cause a series of oral cavity issues [1]. Human gingival fibroblasts (HGFs) locate gingival mesenchymal tissues, and play an important role in maintain- ing the normal structure of gingival and remod- eling process. Moreover, the long-term stability of dental implant is closely associated with HGFs [2]. Some studies showed that nicotine could inhibit HGFs proliferation in a dose- dependent manner. Cigarette extracts can cause abnormal structure of HGFs, cellular DNA injury and facilitate HGFs apoptosis. Tobacco can also cause breakage of double stranded DNA in oral fibroblast [3, 4]. Low con- centration of cotinine and nicotine can de- crease the adhesion of HGFs precursors on dental slices, whilst high dosage of cotinine

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Effects of platelet rich plasma (PRP) on human gingival fibroblast, osteoblast and periodontal ligament cell behaviour

Effects of platelet rich plasma (PRP) on human gingival fibroblast, osteoblast and periodontal ligament cell behaviour

The concentrated growth factors released from PRP was then investigated on the migration and proliferation of various cell-types from the oral cavity (Figs. 2, 3 and 4). Interestingly, it was found that gingival fibroblasts dem- onstrated the ability to further promote cell migration and proliferation in response to PRP when compared to PDL cells and osteoblasts. Furthermore, COL1 staining was also more significantly upregulated in gingival fibro- blasts when compared to PDL cells and osteoblasts. Interestingly, it was observed that PRP had no effect on the ALP activity of PDL cells and actually significantly down-regulated in vitro mineralization potential of PDL cells by demonstrating lower levels of alizarin red staining (Fig. 4) whereas its effect on osteoblasts demonstrated

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Original Article The inhibitory effect of mesenchymal stem cell conditioned medium on the secretion of inflammatory factor of human gingival fibroblast induced by lipopolysaccharide

Original Article The inhibitory effect of mesenchymal stem cell conditioned medium on the secretion of inflammatory factor of human gingival fibroblast induced by lipopolysaccharide

MSC conditioned medium has been widely applied in the clinical therapy. For instance, the medium accelerates wound healing with fewer scars [26]. Besides, it also attenuated the degeneration of axons and myelin of spinal motor neurons [27]. Mesenchymal stem cells played anti-inflammatory roles during innate immune responses [28]. Recent studies also found that MSCs conditioned medium can pro- mote human mesenchymal stem cell growth, proliferation, and differentiation potential [29]. However, there is still lack of reports about the role of MSCs conditioned medium on HGFs induced by LPS. This study showed that MSCs conditioned medium treatment significantly elevated cell proliferation, declined cell ap- optosis, attenuated Caspase 3 activity, and decreased IL-8, TNF-α, and PGE2 levels, which was consistent with previous finding that the expressions of TNF-α and IL-6 in macrophages were suppressed by MSCs conditioned medium [28]. MSCs conditioned medium exerted thera- peutical effect on liver injury by inhibiting inflammation [30]. In a similar fashion, our data unravel that, by inhibiting inflammatory cyto- kines secretion, MSCs conditioned medium can protect HGFs from LPS induction and induces cell apoptosis, although the specific mechanism remains to be further discussed. Inflammatory responses in the gingival tissues were demonstrated to be associated to patients with periodontitis [31]. This study illustrates that MSCs conditioned medium plays an inhibi- tory role during the development of periodonti- tis via the alleviation of inflammatory respons- es, which lays academic basis for further treatment of periodontitis.

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Original Article Effect of mesenchymal stem cell conditioned medium on human gingival fibroblast proliferation and collagen synthesis

Original Article Effect of mesenchymal stem cell conditioned medium on human gingival fibroblast proliferation and collagen synthesis

HGFs isolation and grouping: The impacted mandibular tooth and accessory gingival tissue were collected from the healthy adult. Inclusion criteria [17]: healthy volunteers without acute or chronic inflammation during tooth extraction. Exclusion criteria [17]: subjects combined with other diseases, such as infectious diseases, malignant tumor, severe liver and kidney dis- ease, pulmonary fibrosis, bone metabolic dis-

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Human and Dogs’ Gingival Stem Cells are Different

Human and Dogs’ Gingival Stem Cells are Different

mesenchymal stem cells. Although previous report has shown that these cells possess some stem cell surface markers [4], these markers need to be studied more in order to confirm or otherwise discard classifying them as potential stem cells. Previous studies have shown that LIPUS has anabolic effects on human gingival fibroblast cells [2]. However, it is not known if LIPUS may or may not change these stem cell surface markers in CGMCs and HGMCs. Hence, it was our study aim to test the effect of LIPUS on HGMCs. Also, in translational research, higher animals like beagle dogs are considered as an ideal animal model for preclinical experiments. Hence, it was also important to know if CGMCs possess stem cell markers or not and is so, what would be the effect of LIPUS on these stem cell surface markers. Our study evaluated for the first time the mesenchymal stem cell markers in CGMCs and the effect of LIPUS application on these markers as well as cell viability (as evaluated by MTT) and ALP, DNA activities/concentration. Cell viability assay showed that LIPUS increased MTT absorbance in both HGMCs and CGMCs. This reflects that LIPUS did not show any deleterious effect on either cell types. This is in agreement with previous studies [2,11-15,19,26,27]. Our study showed that beagle dogs’ gingival mesenchymal cells lack the expression of CD73 and CD 105. Previous position paper reported that in order to identify cells as mesenchymal stromal cells and have pluripotent characteristics, these cells must be expressing CD73, CD90 and CD105 more that 95%. The lack of expression of CD73, CD105 by CGMSCs suggest that CGMCs cells cannot be identified or considered as potential stem cells as they fail to have these important criteria. In addition, CGMCs expression of CD 90 was 50% without LIPUS and 45% with LIPUS. Regardless the decreased CD90 expression by LIPUS in CGMCs, these cells at the current time may not be considered as stem cells. Also, HGMCs

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Mass Spectrometry-Based Proteomics Analysis Of Bioactive Proteins In EMD That Modulate Adhesion Of Gingival Fibroblast To Improve Bio-Integration Of Dental Implants

Mass Spectrometry-Based Proteomics Analysis Of Bioactive Proteins In EMD That Modulate Adhesion Of Gingival Fibroblast To Improve Bio-Integration Of Dental Implants

model in combination with mass spectrometry to investigate surface specificity for protein binding of three different Ti surfaces (PT, SLA, and SLActive). By applying this approach, we showed that the Ti surfaces had a low specificity for protein binding due to high similarity on the pellicle composition despite differences in characteristics between surfaces. The lack of binding specificity led us to explore the EMD composition utilizing another strategy. Through the MudPIT methodology, we fractionated EMD in 32 fractions to characterize its proteome. We identified 2000 proteins through tandem mass spectrometry (MS/MS) including novel proteins that are associated with EMD biological activity, i.e. biomineralization, wound healing and biological adhesion. The obtained EMD fractions were then applied to human gingival fibroblast (HGF) to evaluate their capability to promote cell adhesion on a coated surface. The adhesion assay indicated that two EMD fraction (F23 and F24), which contained the adhesion proteins fibrillin-1 and tenascin C, showed a significantly higher response than native EMD and other EMD fractions. Overall, the work presented herein indicated the need for more studies on surface-proteins interaction given the low surface specificity presented by each Ti surface. Also, this thesis provided an in-depth insight on the complexity of EMD protein composition, including the identification of novel proteins that are related to EMD biological activity such as the adhesion of gingival fibroblasts.

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Aesthetics in Periodontal Therapy!

Aesthetics in Periodontal Therapy!

Gingival defects due to gingival or periodontal disease such as interdental craters, alteration in the physiologic contour of gingiva-Gingival soft tissue deformities where physiologic contour of gingiva is altered include gingival craters and clefts, gingival enlargement and interdental crater resulting from necrotizing ulcerative periodontitis. Gingivoplasty is a procedure by which gingiva is reshaped to correct deformities. 18 It is similar to gingivectomy but the

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Functional fingerprinting of human mesenchymal stem cells using high-throughput RNAi screening

Functional fingerprinting of human mesenchymal stem cells using high-throughput RNAi screening

Functional fingerprinting based on cell viability might complement current MSC characterization methods, such as surface marker expression and differentiation as- says, to increase the repertoire for a better definition of MSCs. Cell viability can easily be quantified in a high- throughput manner and shows a good perturbation and dynamic range between different conditions and cell types [14]. High-throughput genetic screening ap- proaches are broadly applied in model organisms and cells to delineate cellular pathways and identify com- ponents of cellular networks. RNA interference (RNAi) allows the systematic depletion of RNA transcripts and to analyze the influence of genes on diverse cellular processes. While such methods have been successfully applied in model organisms and in transformed cell lines, their use to dissect processes in primary cells has thus far been limited due to technical challenges. However, recent insights into genotype-to-phenotype relationships of complex traits in primary cells, such as MSCs, will allow an in-depth understanding of the functional properties of these cells. By adapting high- throughput methods originally developed for transformed cell lines, we have come up with strategies to quantita- tively characterize and define the phenotype of human cell preparations [15–17]. Transferring established screening protocols from immortalized cell lines to primary cells is not trivial and technical bottlenecks include required cell numbers, susceptibility to small interfering RNA (siRNA) transfection, assay robustness and the issue of heterogen- eity accompanying primary cells [17].

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Original Article Lactose induced redox-dependent senescence and activated Nrf2 pathway

Original Article Lactose induced redox-dependent senescence and activated Nrf2 pathway

Lactose is a disaccharide that is composed of galactose and glucose and is originally found in milk and milk products. It is therefore one of the most common nutrients taken by human beings as daily dietary intake. Upon absorption, it is hydrolyzed by lactase to glucose and galac- tose on the surface of the small intestine [13]. However, a deficiency of lactase would prevent its metabolism, leading to lactose intolerance. Approximately 15% of people in Northern Eur- opean are reported as lactose intolerant, this percentage is even higher in other races, up to 80% in Latinos and nearly 100% in Chinese [14]. People with lactose intolerance are unable to digest it completely, leading to accumulation of lactose in the small intestine that is reflect- ed by an array of intestinal distress [13]. How- ever, effects of accumulated lactose on cellular senescence and aging remain largely unknown. In the present study, we have studied the ef- fects of lactose on the senescence of human fetal lung fibroblast cell lines (MRC-5 cell lines), a model for research on aging. We found la- ctose can increase the expression of SA-β-gal [15] and cyclin-dependent kinase (CDK) inhi- bitor p16 ink4a , which are the most widely used

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In vitro study of RRS HA injectable mesotherapy/biorevitalization product on human skin fibroblasts and its clinical utilization

In vitro study of RRS HA injectable mesotherapy/biorevitalization product on human skin fibroblasts and its clinical utilization

Abstract: Mesotherapy/biorevitalization with hyaluronic acid (HA) is a treatment approach currently used for skin rejuvenation. Various products with a wide range of polycomponent formulations are available on the market. Most of these formulations contain noncross-linked HA in combination with a biorevitalization cocktail, formed by various amounts of vitamins, minerals, amino acids, nucleotides, coenzymes, and antioxidants. Although ingredients are very similar among the different products, in vitro and clinical effects may vary substantially. There is a real need for better characterization of these products in terms of their action on human skin or in vitro skin models. In this study, we analyzed the effect of the RRS ® (Repairs, Refills,

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Morphological and functional characteristics of human gingival junctional epithelium

Morphological and functional characteristics of human gingival junctional epithelium

Cytokeratins (CKs) are intermediate filament proteins of cytoskeleton family and are the major structural pro- teins in epithelial cells. As we know, the expression of keratins is one of the definitive characteristics of epithe- lial cells and reflects the biological properties of epithe- lial cells, including their origination, development, histological type, and level of differentiation [17,18]. Sev- eral researches have studied the expression and distribu- tion of a variety of CKs (CK-pan, 5/6, 7, 8/18, 10/13, 16, 17, 19, 20) in periodontal tissues of humans and animals, and the expression of some keratins in gingival epithe- lium were determined [15,19-21]. For example, the ex- pression patterns of CK10/13, 16, 19 in JE were different from that in OGE and SE; The especially high expression of CK19 in all layers of JE made it became a characteris- tical histological marker for JE in vivo [3,22-24]. How- ever, the expressions of various types of cytokeratin in JE and the difference with OGE and SE have not been sys- tematically reported.

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Membrane cofactor protein of complement is present on human fibroblast, epithelial, and endothelial cells

Membrane cofactor protein of complement is present on human fibroblast, epithelial, and endothelial cells

Membrane cofactor protein (MCP) of the complement system is a iC3/C3b binding molecule with cofactor activity that has been identified on all human peripheral blood cells except erythrocytes. Human mononuclear and platelet MCP is dimeric with molecular weights of 68,000 and 63,000 and is expressed in three phenotypic patterns. To further determine its tissue distribution, surface-labeled human fibroblast, epithelial, and endothelial cells and cell lines were assessed for the presence of MCP by iC3 affinity chromatography and by immunoprecipitation with a monospecific anti-MCP rabbit polyclonal antibody. All sources of adult and fetal fibroblast and epithelial cells and cell lines examined and umbilical vein endothelial cells expressed MCP. The molecular weight and phenotypic patterns of MCP were similar to those of peripheral blood cells. MCP was synthesized by fibroblast and epithelial cell lines. Solubilized extracts of these cell lines expressed factor I-dependent cofactor activity for the first cleavage of iC3/C3b which was abrogated by removal of MCP. Expression of MCP was modulated by SV40 transformation of two fetal fibroblast lines. There was a 5- to 10-fold increase in expression of MCP and a preferential expression of the lower species such that the phenotypic designation was changed. The wide tissue distribution and activity profile of MCP suggest that it is likely to play an important role in the regulation of the complement […]

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Fibroblast heterogeneity: implications for human disease

Fibroblast heterogeneity: implications for human disease

The mammalian dermis represents an archetypal mesenchymal tissue largely com- posed of ECM elements, primarily type I and type III collagens, but also other collagen subtypes, proteoglycans, and elastin (4). It serves as a structural scaffold, and a base- ment membrane enriched in type IV colla- gen (44) separates it from the epidermis, a stratified squamous epithelium that forms the outermost layer of the skin. The dermis is penetrated by numerous appendageal structures, including hair follicles and sweat glands, which manifest as specialized invag- inations of the epithelium, for which the basement membrane is in continuity with the interfollicular epidermis (Figure 1). The detailed microstructure of the dermis differs between human and mouse skin. In humans, there is a relatively low density of hair folli- cles on body sites outside of the scalp, with intervening interfollicular epidermis exhib- iting invaginations known as rete ridges. In mice, the density of hair is higher and rete ridges are not prominent (Figure 2).

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Evaluation of Salivary Leptin Levels in Healthy Subjects and Patients with Advanced Periodontitis

Evaluation of Salivary Leptin Levels in Healthy Subjects and Patients with Advanced Periodontitis

Purwar et al, [29] in 2015 evaluated leptin levels in saliva and serum of patients with chronic periodontitis and healthy controls. They conclu- ded that the level of salivary leptin in chronic periodontitis patients was significantly lower than that in healthy controls, consistent with the results of the current study. They also noticed that the level of serum leptin increased in patients with periodontitis compared to healthy volunteers. In another study in 2015, Purwar et al, [30] found that appropriate non-surgical therapy can increase the leptin levels in the saliva of chronic periodontitis patients. Selvarajan et al, [31] in 2015 examined and compared the GCF concentration of leptin in patients with perio- dontal disease and healthy subjects. They observed a substantial decrease in leptin levels in GCF as the periodontal disease progressed. The authors suggested a potential protective role for leptin with regard to periodontal health. However, with the progression of periodontal disease, the protective effects of leptin on gingival tissues decrease due to a decrease in its concentration. Karthikeyon and Pradeep [16] in 2007 reported a significant decrease in the leptin concentrations of GCF with an increase in periodontal tissue destruction, with the highest and lowest leptin concentrations in the GCF in healthy subjects (2658 pg/mL) and patients with chronic periodontitis (1312 pg/mL), respectively. In their study, GCF samples were evaluated,

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Effect of novobiocin on the viability of human gingival fibroblasts (HGF-1)

Effect of novobiocin on the viability of human gingival fibroblasts (HGF-1)

Viability assays in gingival fibroblasts, HGF-1 employed the fluorescence test of Live⁄Dead Viability⁄Cytotoxicity Kit (Invitrogen, USA). The test allows to distinguish vi- able cells (stained with green-fluorescent calcein-AM) from dead cells (stained with red-fluorescent ethidium homodimer-1). In the studies novobiocin (Sigma-Aldrich) was used. The culture medium consisted of DMEM (ATCC) enriched with 10% FBS (Sigma-Aldrich). The stu- dies took advantage of 24 h cultures of gingival fibroblasts, HGF-1, which following incubation were subjected to triple rinsing. The tests in triple repetitions were con- ducted in culture Lab-Tek Chamber Slides (Nunc) in pres- ence of culture medium alone - control (0.5 × 10 6 cells of HGF-1) and in presence of novobiocin (in concentrations of 0.1, 0.5, 1, 2.5 or 5 mM/L/0.5 × 10 6 HGF-1 cells). The samples were incubated for 20 h at 37°C in presence of 5% CO 2 . In addition, the samples were incubated with

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Phytochemical analysis and antiproliferative studies of various extracts of mollugo cerviana

Phytochemical analysis and antiproliferative studies of various extracts of mollugo cerviana

In conclusion, the results obtained from short term cytotoxicity study and antiproliferative activity reveals that various extracts of Mollugo cerviana have significant cytotoxic activity against all the animal and human cancer cell lines studied. Further studies in our laboratory are in progress for the isolation and identification of phytochemical compounds and to ensure the medicinal properties of the plant in vivo correlate with its anticancer activity.

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Plant protein hydrolysates as a supplement for medium of human skin fibroblast 1184 cell culture

Plant protein hydrolysates as a supplement for medium of human skin fibroblast 1184 cell culture

Medium adalah bahan yang paling penting dalam pengkulturan sel. Setiap medium terdiri daripada dua bahagian yang utama iaitu medium asas dan serum. Sumber utama serum ialah daripada darah haiwan. Serum daripada sumber haiwan mempunyai banyak kekerangan termasuklah dicemari virus. Medium tanpa serum merupakan alternatif terbaik untuk menyelesaikan masalah ini. Namun medium tanpa serum tidak mencukupi untuk pertumbuhan sel. Oleh sebab itu, beberapa nutrien tambahan perlu dimasukkan di dalam medium tanpa serum. Salah satu sumber nutrien tambahan adalah daripada tumbuh-tumbuhan yang mungkin dapat mengurangkan kos. Dalan kajian ini protein hidrolisat dari beberapa tumbuhan berlainan seperti soya, bijan, lidah buaya, beras dan gandum telah dikaji terhadap sel fibroblast manusia (HSF 1184). Kesemua protein hidrolisat dihasilkan dengan kaedah hidrolisis protein menggunakan pelbagai enzim komersil dari sumber bukan haiwan. Protein hidrolisat dikategorikan mengikut ciri-ciri keterlarutan dan saiz peptida. Pertumbuhan HSF 1184 adalah berbeza dan bergantung kepada jenis hidrolisat yang dicampurkan ke dalam medium DMEM mengandungi FBS dan tanpa FBS. Hidrolisat daripada enzim exopeptidase seperti Flavourzyme memberikan kesan negatif terhadap pertumbuhan sel HSF 1184. Manakala hidrolisat dari enzim endopeptidase diperlukan untuk pertumbuhan HSF 1184. Oleh kerana protein tumbuhan

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Gingival enlargement, an over-exuberant response to a range of local and systemic conditions, is a common finding in a routine dental practice. A greater part of such enlargements, however, occurs as an unwanted side effect of medications used mainly for systemic treatment for which the gingival tissue is not the projected target organ. An increasing number of medications are recently associated with gi ngival enlargement. Among those medications, gingival enlargement induced by nifedipine has been extensively reported. Although frequently reported, nifedipine -induced gingival enlargement remains a perpetual topic of discussion due to its several obscure features. This review summarizes the importance of gingival overgrowth as a clinical problem following systemic medication of nifedipine, and highlights recent advances that lead to an enhanced understanding of the biological mechanism underlying the overgrowth. A simple flowchart summari zing the various treatment options have been proposed. We also report a series of s ix patients with this condition.

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Influence of Polycaprolactone and Basic Fibroblast Growth Factor on Gingival Fibroblasts

Influence of Polycaprolactone and Basic Fibroblast Growth Factor on Gingival Fibroblasts

inflammation persists with neutrophil infiltration gradually subsiding as macrophage influx becomes predominant. The phenotype of the macrophages switches to promote repair through the release of growth factors such as transforming growth factor-β1 (TGF- β1) which enables the transition to development of granulation tissue and the proliferative stage of repair (Leibovich and Ross 1975, Wikesjo and Selvi 1999). As granulation tissue forms up to day seven following injury, fibroblasts and endothelial cells migrate into the wound area and proliferate, a provisional matrix is deposited, and angiogenesis occurs (Wikesjo and Selvi 1999, Polimeni, Xiropaidis et al. 2006). Fibroblast migration into the granulation tissue requires downregulation of integrins for collagen attachment (α2β1) and upregulation of integrins for interaction with components of the provisional matrix: fibrin (αVβ3), fibronectin (α4β1) and vitronectin (αVβ5) (Aukhil 2000). Following the events in the initial seven days post-injury, the wound enters the matrix remodeling phase wherein the granulation tissue matrix is remodeled into a more collagen rich, mature and stable matrix followed by the appearance of myofibroblasts which contract the wound (Wikesjo and Selvi 1999, Polimeni, Xiropaidis et al. 2006). Although the initial steps of wound healing are essentially analogous to most soft tissue healing, proper periodontal healing requires the connective tissue to make adequate adhesion to the tooth root through anchorage of collagen fibers to the cementum (Wikesjo and Selvi 1999).

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