The future for the application of fibroblast growth factor 2 in modern wound healing

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00159816%3A_____%2F23%3A00077934" target="_blank" >RIV/00159816:_____/23:00077934 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00216305:26620/22:PU146795 RIV/65269705:_____/23:00077934 RIV/00216224:14110/23:00134621

Výsledek na webu

<a href="https://reader.elsevier.com/reader/sd/pii/S030541792200273X?token=6C9EDDF6711ACC81DD0A8F88E6C90BDC01E534763E5D8C1170D5B3DBDAA7D389DC1DDFBFFC275DB61849C05BC0ACA7D9&originRegion=eu-west-1&originCreation=20230418080512" target="_blank" >https://reader.elsevier.com/reader/sd/pii/S030541792200273X?token=6C9EDDF6711ACC81DD0A8F88E6C90BDC01E534763E5D8C1170D5B3DBDAA7D389DC1DDFBFFC275DB61849C05BC0ACA7D9&originRegion=eu-west-1&originCreation=20230418080512</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.burns.2022.10.007" target="_blank" >10.1016/j.burns.2022.10.007</a>

Jazyk výsledku

angličtina

Název v původním jazyce

The future for the application of fibroblast growth factor 2 in modern wound healing

Popis výsledku v původním jazyce

Representatives of the fibroblast growth factor (FGF) family regulate a wide range of biological functions important for every phase of wound healing, such as cell proliferation, migration, and differentiation. The name of the family is derived from the primary assumption that FGF proteins (FGF1 and FGF2) are important for promoting fibroblast migration and proliferation. Today, FGFs are known to intervene in a wide range of biological functions of various cell populations. Their function is mediated by binding to specific receptors (tyrosine-kinase receptors), which leads to their activation and transmembrane signal transduction into the cell [1].Based on sequence similarities, biochemical functions and their own activity, the individual representatives of the FGF family were divided into seven subgroups (subfamilies). The FGF1 subfamily (composed of FGF1 and FGF2) plays a key role in the healing and regeneration processes. Damaged macrophages and endothelial cells at the site of injury release FGF2 and FGF1, thereby directly influencing the healing process; FGF2 then plays a key role also in the scarless healing process, which makes it potentially useful in regenerative medicine) [2].Human recombinant FGF2 (otherwise known as FGF2 or bFGF) was first used in a clinical application in Japan. [3]. Many studies showed that FGF2 shortened the healing time of wounds and improved the quality of healing, accelerated the re-epithelialization, improved the appearance of scar tissue, prevented contractures, and protected against microbial proliferation [2].During the past decade, numerous publications focused on clinical testing of FGF2 in the treatment of skin defects. Several studies have successfully translated the results into clinical practice. In 2000, recombinant human FGF2 was approved by the Chinese Food and Drug Administration for the treatment of chronic wounds, including chronic granulating wounds, ulcers, bedsores, traumatic and surgical wounds and burn wounds, without apparent adverse effects [4]. Based on clinical research and good safety data, the Clinical Practice Guidelines for Burn Injuries published by the Japanese Society for Burn Injuries in 2009 recommended FGF2 as a treatment for second-degree burns [4]. Most recently, the extensive review by Gragnani et al. (2022) showed that FGF2 can be effective in treating injuries caused by burns without generating adverse effects. Unfortunately, the use of FGF2 in routine clinical practice has not yet been widely implemented [5].The described results seem to be very promising indicators for the way forward, showing the potential of FGF2 in promoting the repair of skin damage caused by various etiologies. However, the FGF2 application is limited by the cost of recombinant protein production and by the need for repeated application resulting from its low stability both in vivo and in vitro [6]. In a study by Dvorak et al. (2018), this problem was overcome by using a generalizable computer-aided protein engineering approach. The combination of computer-assisted &quot;bioengineering&quot; with a unique functional screening of novel variants led to the creation of a thermally stable FGF2 variant with unique stability and uncompromised biological function. FGF2-STAB(R)(Dvorak et al., 2018, European Patent EP3380508B1), carrying nine amino acid substitutions, is a stabilized growth factor that retains full biological activity even after 20 days at 37 oC, facilitating new and more efficient growth of FGF2-dependent cell cultures, with fewer media exchanges. FGF2-STAB(R) is fully biologically active, as demonstrated by its ability to engage the FGF receptor downstream signaling pathways, maintain human embryonic stem cells in the undifferentiated state, and inhibit chondrocyte growth. The existence of FGF2-STAB(R) brings a whole new range of application possibilities and potential clinical use not only in the case of topical application but also in the case of chemical incorporation of the protein into other biomaterials. The successful demonstration of rapid protein stabilization highlights the power of applied rational protein engineering strategy and should encourage wider use of the described workflow for the stabilization of other growth factors and protein therapeutics that are currently being tested for use in cancer treatment, regenerative medicine, or in a number of metabolism-associated disorders [7].In our recently published paper, Vojtova et al. (2021) described collagen-based scaffolds modified with antibacterial chitosan fibers and the FGF2-STAB(R) protein. The main goal was to prove the safety of the applied stabilized FGF2-STAB(R) protein as well as to demonstrate its efficacy in healing and neovascularization of full-thickness skin regeneration in a rabbit animal model. The biological activity of FGF2 itself has been demonstrated in a number of in vitro tests. Testing of FGF2-STAB(R) on chicken embryos, mouse fibroblasts and B cell lymphocytes demonstrated an increase in the positive effect on angiogenesis compared to wild-type FGF2 while showing zero toxicity on cell cultures. Finally, an in vivo study on a rabbit animal model revealed the superiority of the scaffold with FGF2-STAB(R) in terms of scaffold bioresorption, minimal inflammation, and excellent granulation during skin tissue reconstruction. The results of this work represent the base for further application of FGF2-STAB(R) for biomedical and clinical applications in the fields of tissue engineering and regenerative medicine [8]. Moreover, the pro-healing FGF2-STAB(R) can be further combined with other bioactive compounds, e.g., with antibacterial nanoparticles to treat infected acute or chronic wounds. However, the adsorption of proteins on the nanoparticle surface may influence both the nanoparticles&apos; and proteins&apos; overall bio-reactivity. We found that metabolic activity assays of normal human dermal fibroblasts (NHDF) indicated an antagonistic effect of selenium nanoparticles (SeNPs) and FGF2-STAB(R) at high concentrations of SeNPs. The half-maximal inhibitory concentration (IC50) of SeNPs for NHDF was 18.9 µg/ml and IC80 was 5.6 µg/ml. Based on detailed in vitro testing, the optimal concentrations of additives in the collagen/chitosan scaffolds were determined, specifically 1 µg/ml of FGF2-STAB(R) and 1 µg/ml of SeNPs [9].We are currently working on several randomized controlled trials using FGF2-STAB(R) to utilize this protein in combination with various scaffolds in burn wound management and tissue regeneration and intend to offer it for future commercial/clinical use. We successfully demonstrated the superiority of the observed parameters (clinical, molecular biological, biomechanical) in preliminary in vitro and in vivo tests and now, we are preparing a clinical evaluation with implantable fully resorbable materials as well as with temporary non-resorbable scaffolds.

Název v anglickém jazyce

The future for the application of fibroblast growth factor 2 in modern wound healing

Popis výsledku anglicky

Representatives of the fibroblast growth factor (FGF) family regulate a wide range of biological functions important for every phase of wound healing, such as cell proliferation, migration, and differentiation. The name of the family is derived from the primary assumption that FGF proteins (FGF1 and FGF2) are important for promoting fibroblast migration and proliferation. Today, FGFs are known to intervene in a wide range of biological functions of various cell populations. Their function is mediated by binding to specific receptors (tyrosine-kinase receptors), which leads to their activation and transmembrane signal transduction into the cell [1].Based on sequence similarities, biochemical functions and their own activity, the individual representatives of the FGF family were divided into seven subgroups (subfamilies). The FGF1 subfamily (composed of FGF1 and FGF2) plays a key role in the healing and regeneration processes. Damaged macrophages and endothelial cells at the site of injury release FGF2 and FGF1, thereby directly influencing the healing process; FGF2 then plays a key role also in the scarless healing process, which makes it potentially useful in regenerative medicine) [2].Human recombinant FGF2 (otherwise known as FGF2 or bFGF) was first used in a clinical application in Japan. [3]. Many studies showed that FGF2 shortened the healing time of wounds and improved the quality of healing, accelerated the re-epithelialization, improved the appearance of scar tissue, prevented contractures, and protected against microbial proliferation [2].During the past decade, numerous publications focused on clinical testing of FGF2 in the treatment of skin defects. Several studies have successfully translated the results into clinical practice. In 2000, recombinant human FGF2 was approved by the Chinese Food and Drug Administration for the treatment of chronic wounds, including chronic granulating wounds, ulcers, bedsores, traumatic and surgical wounds and burn wounds, without apparent adverse effects [4]. Based on clinical research and good safety data, the Clinical Practice Guidelines for Burn Injuries published by the Japanese Society for Burn Injuries in 2009 recommended FGF2 as a treatment for second-degree burns [4]. Most recently, the extensive review by Gragnani et al. (2022) showed that FGF2 can be effective in treating injuries caused by burns without generating adverse effects. Unfortunately, the use of FGF2 in routine clinical practice has not yet been widely implemented [5].The described results seem to be very promising indicators for the way forward, showing the potential of FGF2 in promoting the repair of skin damage caused by various etiologies. However, the FGF2 application is limited by the cost of recombinant protein production and by the need for repeated application resulting from its low stability both in vivo and in vitro [6]. In a study by Dvorak et al. (2018), this problem was overcome by using a generalizable computer-aided protein engineering approach. The combination of computer-assisted &quot;bioengineering&quot; with a unique functional screening of novel variants led to the creation of a thermally stable FGF2 variant with unique stability and uncompromised biological function. FGF2-STAB(R)(Dvorak et al., 2018, European Patent EP3380508B1), carrying nine amino acid substitutions, is a stabilized growth factor that retains full biological activity even after 20 days at 37 oC, facilitating new and more efficient growth of FGF2-dependent cell cultures, with fewer media exchanges. FGF2-STAB(R) is fully biologically active, as demonstrated by its ability to engage the FGF receptor downstream signaling pathways, maintain human embryonic stem cells in the undifferentiated state, and inhibit chondrocyte growth. The existence of FGF2-STAB(R) brings a whole new range of application possibilities and potential clinical use not only in the case of topical application but also in the case of chemical incorporation of the protein into other biomaterials. The successful demonstration of rapid protein stabilization highlights the power of applied rational protein engineering strategy and should encourage wider use of the described workflow for the stabilization of other growth factors and protein therapeutics that are currently being tested for use in cancer treatment, regenerative medicine, or in a number of metabolism-associated disorders [7].In our recently published paper, Vojtova et al. (2021) described collagen-based scaffolds modified with antibacterial chitosan fibers and the FGF2-STAB(R) protein. The main goal was to prove the safety of the applied stabilized FGF2-STAB(R) protein as well as to demonstrate its efficacy in healing and neovascularization of full-thickness skin regeneration in a rabbit animal model. The biological activity of FGF2 itself has been demonstrated in a number of in vitro tests. Testing of FGF2-STAB(R) on chicken embryos, mouse fibroblasts and B cell lymphocytes demonstrated an increase in the positive effect on angiogenesis compared to wild-type FGF2 while showing zero toxicity on cell cultures. Finally, an in vivo study on a rabbit animal model revealed the superiority of the scaffold with FGF2-STAB(R) in terms of scaffold bioresorption, minimal inflammation, and excellent granulation during skin tissue reconstruction. The results of this work represent the base for further application of FGF2-STAB(R) for biomedical and clinical applications in the fields of tissue engineering and regenerative medicine [8]. Moreover, the pro-healing FGF2-STAB(R) can be further combined with other bioactive compounds, e.g., with antibacterial nanoparticles to treat infected acute or chronic wounds. However, the adsorption of proteins on the nanoparticle surface may influence both the nanoparticles&apos; and proteins&apos; overall bio-reactivity. We found that metabolic activity assays of normal human dermal fibroblasts (NHDF) indicated an antagonistic effect of selenium nanoparticles (SeNPs) and FGF2-STAB(R) at high concentrations of SeNPs. The half-maximal inhibitory concentration (IC50) of SeNPs for NHDF was 18.9 µg/ml and IC80 was 5.6 µg/ml. Based on detailed in vitro testing, the optimal concentrations of additives in the collagen/chitosan scaffolds were determined, specifically 1 µg/ml of FGF2-STAB(R) and 1 µg/ml of SeNPs [9].We are currently working on several randomized controlled trials using FGF2-STAB(R) to utilize this protein in combination with various scaffolds in burn wound management and tissue regeneration and intend to offer it for future commercial/clinical use. We successfully demonstrated the superiority of the observed parameters (clinical, molecular biological, biomechanical) in preliminary in vitro and in vivo tests and now, we are preparing a clinical evaluation with implantable fully resorbable materials as well as with temporary non-resorbable scaffolds.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

30221 - Critical care medicine and Emergency medicine

Projekt

<a href="/cs/project/NU22-08-00454" target="_blank" >NU22-08-00454: Biomimetický vícevrstevný buněčný nosič čtvrté generace pro jednokrokovou úplnou náhradu kůže: z laboratoře do klinické aplikace</a><br>

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2023

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Burns

ISSN

0305-4179

e-ISSN

1879-1409

Svazek periodika

49

Číslo periodika v rámci svazku

2

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

3

Strana od-do

484-486

Kód UT WoS článku

000954799500001

EID výsledku v databázi Scopus

2-s2.0-85149758007