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microRNA Archives | Hair Loss Cure 2020 A number of people asked me to write about microRNA (miRNA) and hair growth last year. The publication of a new study in 2021 now makes this subject more interesting to cover.
MicroRNA and Hair Growth Last year I briefly mentioned a new study from scientists at North Carolina State University pertaining to dermal exosomes and microRNA (miRNA). Per the findings, microRNA shows promise for hair regrowth. In particular, the scientists also studied microRNAs (miRNAs) in dermal exosomes from both the 2D and 3D dermal papilla cells .
The research team identified a particular miRNA called miR-218-5p as a key promoter of hair growth. Per team lead Dr. Ke Cheng, cell therapy with 3D cells based on this work could be an effective treatment for baldness. However, you have to grow, expand, preserve and inject such cells into the balding areas.
“MiRNAs, on the other hand, can be utilized in small molecule-based drugs. So potentially you could create a cream or lotion that has a similar effect with many fewer problems.”
In the latest January 2021 issue of Experimental and Molecular Pathology , a new study from China found yet another type of miRNA that impacts hair loss. In particular, the abnormal expression of miR-133b may inactivate the Wnt/β-catenin pathway. This ultimately impacts hair growth patterns. Moreover, the androgen dihydrotestosterone (DHT) can regulate miR-133b expression. mRNA versus miRNA Note that although closely related, messenger RNA (mRNA) is different from microRNA (miRNA).
Messenger RNA is a single-stranded molecule of ribonucleic acid (RNA) that corresponds to the genetic sequence of a gene. MicroRNA is a small non-coding RNA molecule that functions in RNA silencing and post-transcriptional regulation of gene expression.
Also note the differences between RNA and DNA .
Faster mRNA based Hair Loss Research During the past year, two mRNA based Covid-19 vaccines (from Pfizer-BioNTech and Moderna) have been developed in record time.
Moreover, both groundbreaking vaccines have already been used in millions of humans worldwide as of January 2021. This is the first time in history that mRNA based vaccines have been used in human beings.
A lot of hair loss sufferers (including myself) are excited about the impact of this large-scale experiment. mRNA based clinical trials and research have witnessed a decade of progress in just one year due to these events and rapid emergency government approvals.
Perhaps all of this will boost mRNA (and miRNA) based clinical trials for hair loss related products and vaccines in the future.
When it comes to hair cloning and tissue engineering, 3D culturing related research is booming. The 3D culturing of cells can occur via scaffold techniques or via scaffold free techniques such as 3D spheroids.
Update: July 28, 2020
Recent Studies on 3D Culturing of Hair Cells Over the past few months, a number of studies have come out in relation to 3D culturing of hair cells (in particular, dermal papilla cells). Below, I list them from most recent to oldest.
July 27, 2020 — A new study from North Carolina State University compared 3D versus 2D cultured dermal papilla cells . The 3D dermal papilla cells in a scaffold performed best in regrowing hair. More interestingly, the scientists also studied microRNAs (miRNAs) in dermal exosomes from both the 2D and 3D DP cells. The team then identified one (miR-218-5p) in particular as a key promoter of hair growth. Per lead study investigator Dr. Ke Cheng , the best part is that MiRNAs can be developed into small molecule-based drugs, including creams. A much easier feat in comparison to cell growth, expansion and injection.
June 16, 2020 — Several people in the comments mentioned a new paper titled: “Generation of human hair follicle organoids in vitro and ex vivo by co-culture of primary human hair matrix keratinocytes and dermal papilla fibroblasts”. One of the co-authors of this paper is Dr. Ralf Paus . This experiment succeeded in human scalp skin and not just in mice (h/t reader “Joe”).
April 28, 2020 — Culturing human hair follicle dermal papilla cells in a 3D self-assembling peptide scaffold . The results of this study suggest a new potential 3D culture platform based on a self-assembling peptide scaffold called RAD16-I. This method successfully created hair follicle dermal papilla cells.
3D Culturing of Hair Follicles and Dermal Papilla Cells. Source: Wiley Online Library. April 15, 2020 — A new paper on reconstructed human skin with working hair follicles. Co-authors include the renowned Dr. Roland Lauster and Dr. Gerd Lindner.
The results section has an interesting part titled “Comparison of cultured neopapillae spheroids with scalp hair dermal papillae”. Several of the images of the 3D cultured hair follicles are shown on the right. Neopapillae spheroids were constructed from expanded self‐aggregating dermal papilla cells.
December 26, 2019 — Tissue engineering strategies for human hair follicle regeneration. This review analyzes the various research approaches being developed to tackle hair follicle bioengineering. Lots of discussion about 3D culturing, various types of scaffolding and dermal papilla trichogenicity. For the scientifically inclined readers, Table 1 is quite useful and I am pasting a small part of it here:
December 13, 2018 — An important paper with Dr. Angela Christiano, Dr. Colin Jahoda and Dr. Etienne Wang as co-authors. They created 3D-printed hair follicle molds using a biomimetic approach . I covered this work in detail in my 2018 post on biomimetic tissue engineering of hair follicles .
October 22, 2013
3D Spheroid Culturing of Dermal Papilla Cells This week seems to be full of interesting developments, but the below news made all the global headlines.
Dr. Angela Christiano (Columbia University — US) and Dr. Colin Jahoda (Durham University — UK) just released their latest findings on hair follicle culturing . Their main discovery involves using a “hanging-drop” method of 3D spheroid culturing of dermal papilla cells. As opposed to a regular 2D petri dish culturing method that had failed in the past.
This new 3D method has shown significant success. However, it is still a years away from being able to be used in humans with consistent and safe results.
Media Coverage For more, see this video with the hair follicle blessed Dr. Christiano . Edit: Per the Fox News video in the link at the bottom, it seems like she wears a wig and suffers from Alopecia Areata.
An audio interview from BBC with Dr. Colin Jahoda
And now some other links to their findings:
Article from BBC
Article from New Scientist
Article from NYtimes
Article with Video from Fox News
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Hair loss news blog. Hair cloning for baldness.