Food-Derived Collagen Peptides, Prolyl-Hydroxyproline (Pro-Hyp), and Hydroxyprolyl-Glycine (Hyp-Gly) Enhance Growth of Primary Cultured Mouse Skin Fibroblast Using Fetal Bovine Serum Free from Hydroxyprolyl Peptide

Asai, Tomoko; Oikawa, Fumi; Yoshikawa, Kazunobu; Inoue, Naoki; Satô, Kenji

doi:10.3390/ijms21010229

Cited by 30 publications

(23 citation statements)

References 21 publications

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“…The latter two compounds are readily separated by the m/z values of the precursor ions. The non‐spiked serum sample clearly shows the high endogenous levels of two of the analytes (B: p and G: P p ), which agrees with the findings by Asai et al 28 Compound H, the tripeptide EVF injection standard, can be used to assess the internal standard signals when unforeseen issues occur during a future analytical run. The internal standards are all present at the same concentration in the internal standard mix and in the end extract (145 ng/mL, expressed as analyte).…”

Section: Resultssupporting

confidence: 89%

Reliable and generic liquid chromatography/mass spectrometry quantification of short peptides using a stable‐isotope‐labeled labeling agent

Kleinnijenhuis¹,

Vergauwen²,

Holthoon³

et al. 2020

Rapid Comm Mass Spectrometry

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Rationale It is important to investigate the behavior of protein hydrolysate components in both in vitro and in vivo studies, to support the elucidation of their biological functions. As protein hydrolysates and biological matrices are highly complex mixtures, it is essential to apply fully reliable and flexible analytical approaches. Methods A novel and generic Liquid Chromatography/Mass Spectrometry methodology was developed to analyze short peptides. A stable‐isotope‐labeled labeling agent 6‐aminoquinolyl‐N‐hydroxysuccinimidyl carbamate (13C3) was synthesized and used to prepare internal standards from non‐labeled analyte peptides. The amino acid and peptides p, pG, Pp, GPp and PpG (where p stands for hydroxyproline) were used for proof of principle. Results The method showed acceptable performance in solvent, in simulated gastrointestinal fluid and in serum. The (linear) dynamic range expanded to over four orders of magnitude, which is very useful when multiple analytes are analyzed in a biological matrix, due to the large differences in concentrations observed for endogenous and protein hydrolysate components. The method provides absolute‐quantitative results and is fully accountable on the single‐sample and single‐component level. Conclusions The methodology can be applied to reliably quantify protein hydrolysate nutraceutical components at various stages during their in vivo processing. Internal standards can also be synthesized for other short peptides whenever they are expected to have biological relevance and require quantification. Overall this provides an excellent analytical tool to support the elucidation of the biological functions of protein hydrolysate components.

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Section: Resultssupporting

confidence: 89%

Reliable and generic liquid chromatography/mass spectrometry quantification of short peptides using a stable‐isotope‐labeled labeling agent

Kleinnijenhuis¹,

Vergauwen²,

Holthoon³

et al. 2020

Rapid Comm Mass Spectrometry

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“…However, daily intakes of food-derived collagen peptides enhance the growth of fibroblasts. Pro-Hyp and Hyp-Gly dipeptides play an important role in the proliferation of fibroblasts [119]. It has been demonstrated that the rest of the fragmented collagen inhibits synthesis of procollagen by fibroblasts, blocks their proliferation, induces a senescent fibroblastic morphology, and stimulates the expression of matrix metalloproteinases (MMP) [120].…”

Section: Enhancement Of Fibroblast Production and Extracellular Matrimentioning

confidence: 99%

Collagen Hydrolysates for Skin Protection: Oral Administration and Topical Formulation

Aguirre-Cruz¹,

León-López

Cruz-Gómez³

et al. 2020

Antioxidants

131

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Antioxidants are molecules that delay or inhibit the oxidation of other molecules. Its use significantly increased in recent years in the diet of people. Natural antioxidants are replacing the use of synthetic antioxidant ingredients due to their safety, nutritional, and therapeutic values. Hydrolyzed collagen (HC) is a popular ingredient considered to be an antioxidant. This low molecular weight protein has been widely utilized due to its excellent biocompatibility, easy biodegradability, and weak antigenicity. It is a safe cosmetic biomaterial with good moisturizing properties on the skin. The antioxidant properties of HC are conditioned to the size of the molecule: the lower the molecular weight of peptides, the greater the ability to donate an electron or hydrogen to stabilize radicals. The antioxidant capacity of HC is mostly due to the presence of hydrophobic amino acids in the peptide. The exact mechanism of peptides acting as antioxidants is not clearly known but some aromatic amino acids and histidine are reported to play an important role in the antioxidant activity. Oral ingestion of HC increases the levels of collagen-derived peptides in the blood torrent and improves the skin properties such as elasticity, skin moisture, and transepidermal water loss. Additionally, daily intakes of HC protect the skin against UV melasma, enhances the fibroblast production and extracellular matrix of the skin. HC has been identified as a safe cosmetic ingredient for topical formulations with good moisturizing properties at the stratum corneum layer of the skin. It reduces the effects of skin aging (dryness, laxity, and wrinkles). The use of HC as a principal ingredient in safe formulations for skin protection was reviewed and compared when it is used by topical and/or oral administration.

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“…In cell culture systems, fibroblasts proliferate on plastic substrate in the presence of fetal bovine serum (FBS), which is rich in protein growth factors and fibronectin. The removal of low molecular weight compounds from FBS does not affect the growth of fibroblasts on the plastic substrate (Asai et al, 2020a ). Fibroblasts have to attach to the substrates for survival.…”

Section: Introductionmentioning

confidence: 99%

“…In cell culture systems using non-coated culture plates, fibroblasts are attached to the plate surface via fibronectin, which is present in FBS and promote cell proliferation (Hayman and Ruoslahti, 1979 ). However, some researchers have used collagen gel-coated plates for the cultivation of fibroblasts (Yoshisato et al, 1985 ; Nishiyama et al, 1989 ; Kono et al, 1990 ; Shigemura et al, 2009 ; Asai et al, 2020a , b ). The collagen solution becomes to be polymerized at neutral pH after incubation at 37°C.…”

Section: Introductionmentioning

confidence: 99%

“…However, rational information is still limited for the fibroblasts under such context. Our group have previously addressed; after total skin excision in mice, collagen di-peptide, prolyl-hydroxyproline (Pro-Hyp), which is generated in the granulation tissue after wounding (Jimi et al, 2017 ) and Pro-Hyp can act as a trigger for growth initiation in the fibroblasts cultured on collagen gel (Asai et al, 2020a , b ; Shigemura et al, 2009 ).…”

Section: Introductionmentioning

confidence: 99%

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Collagen-Derived Di-Peptide, Prolylhydroxyproline (Pro-Hyp): A New Low Molecular Weight Growth-Initiating Factor for Specific Fibroblasts Associated With Wound Healing

Satô

Asai

Jimi

2020

Front. Cell Dev. Biol.

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Many cells and soluble factors are involved in the wound healing process, which can be divided into inflammatory, proliferative, and remodeling phases. Fibroblasts play a crucial role in wound healing, especially during the proliferative phase, and show heterogeneity depending on lineage, tissue distribution, and extent of differentiation. Fibroblasts from tissue stem cells rather than from healthy tissues infiltrate wounds and proliferate. Some fibroblasts in the wound healing site express the mesenchymal stem cell marker, p75NTR. In the cell culture system, fibroblasts attached to collagen fibrils stop growing, even in the presence of protein growth factors, thus mimicking the quiescent nature of fibroblasts in healthy tissues. Fibroblasts in wound healing sites proliferate and are surrounded by collagen fibrils. These facts indicate presence of new growth-initiating factor for fibroblasts attached to collagen fibrils at the wound healing site, where the collagen-derived peptide, prolyl-hydroxyproline (Pro-Hyp), is generated. Pro-Hyp triggers the growth of p75NTR-positive fibroblasts cultured on collagen gel but not p75NTR-negative fibroblasts. Thus, Pro-Hyp is a low molecular weight growth-initiating factor for specific fibroblasts that is involved in the wound healing process. Pro-Hyp is also supplied to tissues by oral administration of gelatin or collagen hydrolysate. Thus, supplementation of gelatin or collagen hydrolysate has therapeutic potential for chronic wounds. Animal studies and human clinical trials have demonstrated that the ingestion of gelatin or collagen hydrolysate enhances the healing of pressure ulcers in animals and humans and improves delayed wound healing in diabetic animals. Therefore, the low molecular weight fibroblast growth-initiating factor, Pro-Hyp, plays a significant role in wound healing and has therapeutic potential for chronic wounds.

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Food-Derived Collagen Peptides, Prolyl-Hydroxyproline (Pro-Hyp), and Hydroxyprolyl-Glycine (Hyp-Gly) Enhance Growth of Primary Cultured Mouse Skin Fibroblast Using Fetal Bovine Serum Free from Hydroxyprolyl Peptide

Cited by 30 publications

References 21 publications

Reliable and generic liquid chromatography/mass spectrometry quantification of short peptides using a stable‐isotope‐labeled labeling agent

Reliable and generic liquid chromatography/mass spectrometry quantification of short peptides using a stable‐isotope‐labeled labeling agent

Collagen Hydrolysates for Skin Protection: Oral Administration and Topical Formulation

Collagen-Derived Di-Peptide, Prolylhydroxyproline (Pro-Hyp): A New Low Molecular Weight Growth-Initiating Factor for Specific Fibroblasts Associated With Wound Healing

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