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Article ; Online: Macromolecular crowding regulates matrix composition and gene expression in human gingival fibroblast cultures.

Ramalingam, Rajesvaran / Jiang, Guoqiao / Larjava, Hannu / Häkkinen, Lari

2023 Volume 13, Issue 1, Page(s) 2047

Abstract: Standard cell cultures are performed in aqueous media with a low macromolecule concentration compared to tissue microenvironment. In macromolecular crowding (MMC) experiments, synthetic polymeric crowders are added into cell culture media to better mimic ...

Abstract	Standard cell cultures are performed in aqueous media with a low macromolecule concentration compared to tissue microenvironment. In macromolecular crowding (MMC) experiments, synthetic polymeric crowders are added into cell culture media to better mimic macromolecule concentrations found in vivo. However, their effect on cultured cells is incompletely understood and appears context-dependent. Here we show using human gingival fibroblasts, a cell type associated with fast and scarless wound healing, that MMC (standard medium supplemented with Ficoll 70/400) potently modulates fibroblast phenotype and extracellular matrix (ECM) composition compared to standard culture media (nMMC) over time. MMC significantly reduced cell numbers, but increased accumulation of collagen I, cellular fibronectin, and tenascin C, while suppressing level of SPARC (Secreted Protein Acidic and Cysteine Rich). Out of the 75 wound healing and ECM related genes studied, MMC significantly modulated expression of 25 genes compared to nMMC condition. MMC also suppressed myofibroblast markers and promoted deposition of basement membrane molecules collagen IV, laminin 1, and expression of LAMB3 (Laminin Subunit Beta 3) gene. In cell-derived matrices produced by a novel decellularization protocol, the altered molecular composition of MMC matrices was replicated. Thus, MMC may improve cell culture models for research and provide novel approaches for regenerative therapy.
MeSH term(s)	Humans ; Collagen Type I/metabolism ; Extracellular Matrix/metabolism ; Cells, Cultured ; Fibroblasts/metabolism ; Macromolecular Substances/metabolism ; Culture Media/pharmacology ; Culture Media/metabolism ; Gene Expression ; Fibronectins/metabolism
Chemical Substances	Collagen Type I ; Macromolecular Substances ; Culture Media ; Fibronectins
Language	English
Publishing date	2023-02-04
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-023-29252-1
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Delayed centrifugation weakens the in vitro biological properties of platelet-rich fibrin membranes.

Robin, Wintermute / Shuichiro, Kobayashi / Leeni, Koivisto / Rana, Tarzemany / Reginaldo, Goncalves / Lari, Häkkinen / Larjava, Hannu

Clinical oral investigations

2024 Volume 28, Issue 4, Page(s) 225

Abstract: Objective: To investigate how delayed blood centrifugation affects the composition of the resultant platelet rich fibrin membrane (PRF, a concentrated growth factor preparation) and its biological effects towards gingival fibroblasts.: Materials and ... ...

Abstract	Objective: To investigate how delayed blood centrifugation affects the composition of the resultant platelet rich fibrin membrane (PRF, a concentrated growth factor preparation) and its biological effects towards gingival fibroblasts. Materials and methods: Blood samples were collected from 18 healthy individuals and centrifuged immediately (T-0), or after a 1-6-minute delay (T-1-6, respectively), to generate PRF. Each PRF membrane was weighed. T-0 and T-6 membranes were incubated for 48 h in cell culture medium at 37 °C to create PRF "releasates" (soluble factors released from the PRF). Human gingival fibroblasts were incubated for 48 h with or without the releasates, followed by RNA isolation and real-time polymerase chain reaction to measure expression of select genes associated with granulation tissue formation, angiogenesis and wound contraction. Additional T-0 and T-6 membranes were used for visualization of leucocyte nuclei and platelets by immunostaining. Results: Immediate centrifugation (T-0) resulted in the largest membranes, T-6 membranes being on average 29% smaller. Leucocytes and platelets were significantly more abundant in T-0 than in T-6 samples. Majority of the fibroblast genes studied were consistently either upregulated or downregulated by the T-0 PRF releasates. However, centrifugation after a 6-minute delay significantly weakened the fibroblast responses. Conclusions: Delayed centrifugation resulted in smaller PRF membranes with fewer leucocytes and platelets and also significantly reduced on the expression of a set of healing-related gingival fibroblast genes. Clinical relevance: The higher expression of wound healing-related genes in gingival fibroblasts by the immediately-centrifuged PRF membranes may increase their biological properties in clinical use.
MeSH term(s)	Humans ; Platelet-Rich Fibrin ; Blood Platelets ; Wound Healing ; Leukocytes ; Centrifugation/methods
Language	English
Publishing date	2024-03-22
Publishing country	Germany
Document type	Journal Article
ZDB-ID	1364490-7
ISSN	1436-3771 ; 1432-6981
ISSN (online)	1436-3771
ISSN	1432-6981
DOI	10.1007/s00784-024-05617-2
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Macromolecular crowding regulates matrix composition and gene expression in human gingival fibroblast cultures

Rajesvaran Ramalingam / Guoqiao Jiang / Hannu Larjava / Lari Häkkinen

Scientific Reports, Vol 13, Iss 1, Pp 1-

2023 Volume 20

Abstract: Abstract Standard cell cultures are performed in aqueous media with a low macromolecule concentration compared to tissue microenvironment. In macromolecular crowding (MMC) experiments, synthetic polymeric crowders are added into cell culture media to ... ...

Abstract	Abstract Standard cell cultures are performed in aqueous media with a low macromolecule concentration compared to tissue microenvironment. In macromolecular crowding (MMC) experiments, synthetic polymeric crowders are added into cell culture media to better mimic macromolecule concentrations found in vivo. However, their effect on cultured cells is incompletely understood and appears context-dependent. Here we show using human gingival fibroblasts, a cell type associated with fast and scarless wound healing, that MMC (standard medium supplemented with Ficoll 70/400) potently modulates fibroblast phenotype and extracellular matrix (ECM) composition compared to standard culture media (nMMC) over time. MMC significantly reduced cell numbers, but increased accumulation of collagen I, cellular fibronectin, and tenascin C, while suppressing level of SPARC (Secreted Protein Acidic and Cysteine Rich). Out of the 75 wound healing and ECM related genes studied, MMC significantly modulated expression of 25 genes compared to nMMC condition. MMC also suppressed myofibroblast markers and promoted deposition of basement membrane molecules collagen IV, laminin 1, and expression of LAMB3 (Laminin Subunit Beta 3) gene. In cell-derived matrices produced by a novel decellularization protocol, the altered molecular composition of MMC matrices was replicated. Thus, MMC may improve cell culture models for research and provide novel approaches for regenerative therapy.
Keywords	Medicine ; R ; Science ; Q
Subject code	571
Language	English
Publishing date	2023-02-01T00:00:00Z
Publisher	Nature Portfolio
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Gingival epithelial cell-derived microvesicles activate mineralization in gingival fibroblasts.

Kobayashi, Shuichiro / Bi, Jiarui / Owen, Gethin / Larjava, Nelli / Koivisto, Leeni / Häkkinen, Lari / Larjava, Hannu

Scientific reports

2022 Volume 12, Issue 1, Page(s) 15779

Abstract: Soft tissue calcification occurs in many parts of the body, including the gingival tissue. Epithelial cell-derived MVs can control many functions in fibroblasts but their role in regulating mineralization has not been explored. We hypothesized that ... ...

Abstract	Soft tissue calcification occurs in many parts of the body, including the gingival tissue. Epithelial cell-derived MVs can control many functions in fibroblasts but their role in regulating mineralization has not been explored. We hypothesized that microvesicles (MVs) derived from gingival epithelial cells could regulate calcification of gingival fibroblast cultures in osteogenic environment. Human gingival fibroblasts (HGFs) were cultured in osteogenic differentiation medium with or without human gingival epithelial cell-derived MV stimulation. Mineralization of the cultures, localization of the MVs and mineral deposits in the HGF cultures were assessed. Gene expression changes associated with MV exposure were analyzed using gene expression profiling and real-time qPCR. Within a week of exposure, epithelial MVs stimulated robust mineralization of HGF cultures that was further enhanced by four weeks. The MVs taken up by the HGF's did not calcify themselves but induced intracellular accumulation of minerals. HGF gene expression profiling after short exposure to MVs demonstrated relative dominance of inflammation-related genes that showed increases in gene expression. In later cultures, OSX, BSP and MMPs were significantly upregulated by the MVs. These results suggest for the first time that epithelial cells maybe associated with the ectopic mineralization process often observed in the soft tissues.
MeSH term(s)	Calcinosis/metabolism ; Cell Differentiation ; Cells, Cultured ; Epithelial Cells/metabolism ; Fibroblasts/metabolism ; Gingiva ; Humans ; Osteogenesis
Language	English
Publishing date	2022-09-22
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-022-19732-1
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Altered composition of the oral microbiome in integrin beta 6-deficient mouse.

Uehara, Osamu / Bi, Jiarui / Zhuang, Deshu / Koivisto, Leeni / Abiko, Yoshihiro / Häkkinen, Lari / Larjava, Hannu

Journal of oral microbiology

2022 Volume 14, Issue 1, Page(s) 2122283

Abstract: In periodontal disease (PD), bacterial biofilms suppress β6 integrin expression transforming growth factor-β1 signaling, resulting in gingival inflammation and bone loss. β6 integrin-null ( ...

Abstract	In periodontal disease (PD), bacterial biofilms suppress β6 integrin expression transforming growth factor-β1 signaling, resulting in gingival inflammation and bone loss. β6 integrin-null (
Language	English
Publishing date	2022-09-12
Publishing country	United States
Document type	Journal Article
ZDB-ID	2523919-3
ISSN	2000-2297
ISSN	2000-2297
DOI	10.1080/20002297.2022.2122283
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Article ; Online: Decontamination of multispecies oral biofilm from rough implant surface by airflow with glycine.

Leung, Kyla / Bi, Jiarui / Giannelis, Georgios / Owen, Gethin / Larjava, Hannu

Clinical and experimental dental research

2021 Volume 8, Issue 1, Page(s) 322–328

Abstract: Objectives: Decontamination of biofilm-colonized rough implant surfaces remains challenging. We investigated the effect of airflow with glycine powder (AFG) on decontamination of mature oral multispecies biofilm from a sandblasted and acid etched (SLA) ... ...

Abstract	Objectives: Decontamination of biofilm-colonized rough implant surfaces remains challenging. We investigated the effect of airflow with glycine powder (AFG) on decontamination of mature oral multispecies biofilm from a sandblasted and acid etched (SLA) titanium surface. Materials and methods: Subgingival dental plaque was cultured on SLA disks anaerobically for 21 days. AFG with various settings and distances was applied directly on the disks with or without previous rinse of 0.9% NaCl. The specimens were then analyzed through scanning electron microscope and remaining bacteria on the implant surface were quantified and statistically compared. Results: Mature oral biofilm with cocci and rods as major morphotypes, as well as spiral- and filamentous-shaped organisms, was formed on the untreated disks. Saline rinsing removed the thick biofilm layer but left numerous of coccoid bacteria in rough surface pits. AFG effectively removed most of the bacteria from the pits. Both 25% and 50% power settings were equally effective at 3-mm distance. With 50% power, AFG successfully removed bacteria at both 3- and 6-mm distance. When AFG was applied on native biofilm without prior rinsing with saline, it effectively removed the biofilm including bacteria in the pits. Conclusion: Application of AFG appears effective in removing bacteria from rough implant surfaces.
MeSH term(s)	Biofilms ; Decontamination ; Dental Implants/microbiology ; Glycine/pharmacology ; Surface Properties
Chemical Substances	Dental Implants ; Glycine (TE7660XO1C)
Language	English
Publishing date	2021-10-26
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2829558-4
ISSN	2057-4347 ; 2057-4347
ISSN (online)	2057-4347
ISSN	2057-4347
DOI	10.1002/cre2.507
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Book ; Thesis: CELLULAR MECHANISMS OF PERIODONTITIS

LARJAVA, HANNU

STUDIES WITH CULTURED HUMAN GINGIVAL FIBROBLASTS AND FETAL RAT CALVARIA EXPOSED TO DENTAL PLAQUE

(PROCEEDINGS OF THE FINNISH DENTAL SOCIETY ; 80 : SUPPL. ; 1)

1984

Series title	PROCEEDINGS OF THE FINNISH DENTAL SOCIETY ; 80 : SUPPL. ; 1 Proceedings of the Finnish Dental Society
Collection	Proceedings of the Finnish Dental Society
Keywords	PERIODONTITIS
Size	59 S.
Publisher	FINNISH DENTAL SOC
Publishing place	HELSINKI
Document type	Book ; Thesis
Thesis / German Habilitation thesis	TURKU, UNIV., DISS., 1984
HBZ-ID	HT002496871
ISBN	951-9401-40-7 ; 978-951-9401-40-9
Database	Catalogue ZB MED Medicine, Health

In stock of ZB MED Cologne/Königswinter

Shelf mark	Loan status	Location
Zs B 1159 -80,Suppl.1/5-	verfügbar in Königswinter	Königswinter
85 E 192	order for loan	Magazin

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Article ; Online: Spray-dried microparticles of encapsulated gefitinib for slow-release localized treatment of periodontal disease.

Baldelli, Alberto / Koivisto, Leeni / Oguzlu, Hale / Guo, Yigong / Häkkinen, Lari / Pratap-Singh, Anubhav / Larjava, Hannu

International journal of pharmaceutics

2023 Volume 642, Page(s) 123137

Abstract: Periodontal disease (PD) can be prevented by local or systemic application of epidermal growth factor receptor inhibitors (EGFRIs) that stabilize αvβ6 integrin levels in the periodontal tissue, leading to an increase in the expression of anti- ... ...

Abstract	Periodontal disease (PD) can be prevented by local or systemic application of epidermal growth factor receptor inhibitors (EGFRIs) that stabilize αvβ6 integrin levels in the periodontal tissue, leading to an increase in the expression of anti-inflammatory cytokines, such as transforming growth factor-β1. Systemic EGFRIs have side effects and, therefore, local treatment of PD applied into the periodontal pockets would be preferrable. Thus, we have developed slow-release three-layered microparticles of gefitinib, a commercially available EGFRI. A combination of different polymers [cellulose acetate butyrate (CAB), Poly (D, L-lactide-co-glycolide) (PLGA) and ethyl cellulose (EC)] and sugars [D-mannose, D-mannitol and D-(+)-trehalose dihydrate] were used for the encapsulation. The optimal formulation was composed of CAB, EC, PLGA, mannose and gefitinib (0.59, 0.24, 0.09, 1, and 0.005 mg/ml, respectively; labeled CEP-gef), and created microparticles of 5.7 ± 2.3 µm in diameter, encapsulation efficiency of 99.98%, and a release rate of more than 300 h. A suspension of this microparticle formulation blocked EGFR phosphorylation and restored αvβ6 integrin levels in oral epithelial cells, while the respective control microparticles showed no effect.
MeSH term(s)	Humans ; Polylactic Acid-Polyglycolic Acid Copolymer ; Gefitinib ; Polyglycolic Acid ; Lactic Acid ; Periodontal Diseases/drug therapy ; Microspheres ; Particle Size
Chemical Substances	Polylactic Acid-Polyglycolic Acid Copolymer (1SIA8062RS) ; Gefitinib (S65743JHBS) ; Polyglycolic Acid (26009-03-0) ; Lactic Acid (33X04XA5AT)
Language	English
Publishing date	2023-06-24
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	428962-6
ISSN	1873-3476 ; 0378-5173
ISSN (online)	1873-3476
ISSN	0378-5173
DOI	10.1016/j.ijpharm.2023.123137
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Zs.A 1409: Show issues

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Article ; Online: Scanning electron microscopic analysis of adherent bacterial biofilms associated with peri-implantitis.

Chang, Jae W / Bi, Jiarui / Owen, Gethin / Shen, Ya / Haapasalo, Markus / Wiebe, Colin / Tarzemany, Rana / Larjava, Hannu

Clinical and experimental dental research

2023 Volume 9, Issue 4, Page(s) 586–595

Abstract: Objectives: Peri-implantitis (PI) is caused by bacteria in the peri-implant space but the consensus on microbial profile is still lacking. Current microbial sampling of PI lesions has largely focused on analyzing bacterial species that have been shed ... ...

Abstract	Objectives: Peri-implantitis (PI) is caused by bacteria in the peri-implant space but the consensus on microbial profile is still lacking. Current microbial sampling of PI lesions has largely focused on analyzing bacterial species that have been shed from the implant surface and captured in the pocket fluid. The purpose of the present study was to investigate the morphotypes of bacteria in biofilm covering the implant threads and explore whether certain morphotypes were associated with PI. Methods: Fourteen failed implants were removed and instantly processed for scanning electron microscope analysis. The implants were imaged at three equally divided sub-crestal levels of the exposed area. Bacterial morphotypes were identified and quantified by three examiners. Mobility and years in function were correlated to the presence of different morphotypes. Results: The implants demonstrated the presence of variable bacterial morphotypes that did not correlate to disease progression in our study. Some implants were dominated by filaments and others showed the presence of combinations of cocci/rods or spirilles/spirochetes. In general, all implants showed variable morphologic biofilm composition. However, individual implants tended to have similar composition throughout the entire implant. Rods and filaments were dominant morphotypes throughout the surfaces and cocci showed increased presence toward the apical third. There were some differences in the biofilm morphology with mobility and time in function. Conclusions: The profiles of bacterial biofilm morphotypes in failing implants with similar clinical presentations were highly variable. While there were significant differences between implants, similar morphotypes in individual implants were often found throughout the entire surface.
MeSH term(s)	Humans ; Peri-Implantitis ; Microscopy, Electron, Scanning ; Electrons ; Bacteria ; Biofilms
Language	English
Publishing date	2023-05-08
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2829558-4
ISSN	2057-4347 ; 2057-4347
ISSN (online)	2057-4347
ISSN	2057-4347
DOI	10.1002/cre2.741
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Book: Oral wound healing

Larjava, Hannu

cell biology and clinical management

2012

Author's details	edited by Hannu Larjava
MeSH term(s)	Periodontal Diseases/rehabilitation ; Mouth/injuries ; Wound Healing ; Oral Surgical Procedures/rehabilitation
Language	English
Size	xvi, 408 p. :, ill.
Publisher	John Wiley & Sons
Publishing place	Chichester, West Sussex
Document type	Book
ISBN	9780813804811 ; 0813804817
Database	Catalogue of the US National Library of Medicine (NLM)

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