Clinical trial starting using Jahoda's method !

[AntiBag Dave]

Not sure if you need a login to view it but here is a link to one of the figures in the paper I mention above:

http://origin-ars.els-cdn.com/content/image/1-s2.0-S0142961212011088-gr6.jpg

[AntiBag Dave]

Interesting stuff Desmond. Here's another exciting paper published by these guys:

http://www.ncbi.nlm.nih.gov/pubmed/23092862

Basically, they show a potentially automatable method for culturing large numbers of "inductive" human DP spheroids of specified sizes. They go on to transplant different sized spheroids into mice to see what effect spheroid size had on the thickness of new hair fibres and found that the whilst the spheroids do create new hair fibres, they are thinner than normal (~15 instead of 50 um), regardless of spheroid size.

In the discussion they mention the possibility of manipulating signalling pathways (e.g. Wnt) to resolve the issues of hair fibre thickness. Given the paper was published over a year ago they may have solved that problem by now. These guys are seriously close...

[rhysmorgan]

Those 400 people should be 400 of us here in the forums. Lord knows we've made more of an effort to warrant a treatment than many others.

Don't be so eager to jump onto new trials. You might end up with a head full of curly pubes.

[southern]

Desmond to the rescue!

[southern]

As carvo said, we should send an email to Sung-Jan Lin. It would be interesting to get a confirmation of the trial and ask him if they are following the 2013 patent method. If that's the case, a clinical treatment might be closer than we expected (I think Taiwan has a special stem cell aproval process, as Japan)

[Desmond84]

Here's what I could dig out of my text books regarding scaffolds composed of polymers (i.e. EVAL):

Introduction

Scaffolds composed of natural polymers have been essential components of tissue engineering since its inception. Polymers are currently used in a wide range of biomedical applications, including applications in which the polymer remains in intimate contact with cells and tissues for prolonged periods. Many of these polymer materials have been tested for tissue engineering applications as well....

Implanting Polymer Scaffolds

The context of cell-polymer interactions in vivo (after implantation) is inherently complex due to the presence of blood, interstitial fluids and multiple cell types in various activation states.

Almost all implanted polymers induce a unique inflammatory response termed the FOREIGN BODY RESPONSE (FBR). The FBR can be divided into several overlapping phases:

- Non-specific protein adsorption
- Inflammatory cell recruitment of neutrophils and macrophages
- Macrophage fusion to form FOREIGN BODY GIANT CELLS (FBGC)
- Involvement of fibroblasts and endothelial cells

The end result of FBR is the formation of FBGC directly on the polymer surface, and the subsequent encapsulation of the implant by a fibrous capsule that is largely avascular. A number of implantation techniques in rodents and larger animals (typically, rabbits, pigs or sheep) have been adopted for the investigation of cell-polymer interactions. Most notably, short-term studies for the analysis of protein adsorption, inflammatory cell recruitment and adhesion, and macrophage fusion most often employ either:

- Intraperitoneal implantation (IP)
- Subcutaneous (SC) cage-implantation --> also known as the wound chamber model

FBGC can then lead to other responses such as inflammation, fibrosis and angiogenesis. There is still much to learn in this area, but it is clear that both the implant material and the physiology of the implant site are important variables.

a) Inflammation
The implantation of polymers through surgical incision means that an initial component of FBR involves a wound-healing like response and it is reasonable to assume that the early inflammatory response is mediated, at least in part, by wound-derived factors. Analysis of several implantation models has shown neutrophils (early) and monocyte/macrophages (late) to be the primary inflammatory cells involved.

FBGC can cause damage to polymer surfaces through their degradative and phagocytic activities and thus pose a significant obstacle to the successful application of polymer-based biomaterials.

b) Fibrosis and Angiogenesis
Unlike wound-healing, the resolution of polymer- associated inflammatory response is characterised by the excessive deposition of highly organised collagenous matrix and a striking paucity of blood vessels. The collagenous capsule can vary in thickness but usually exceeds 100 mcq. The dense and organised nature of the collagen fibres in the capsule could play a role in limiting blood vessel formation.

c) Myofibroblasts
An additional concern with polymer capsulation is the presence of contractile cells, myofibroblasts, which can cause contraction of collagenous capsule and misshape or damage polymer implants. For example, silicone-based breast implants have been shown to be susceptible to this phenomenon!

__________________________________________________ _____________

SO I guess we need to know if they've conducted in vivo studies to analyse if the body tolerates EVAL or not!
__________________________________________________ _____________

We should also try and find out if EVAL is biodegradable in the body? Here's a bit of information on biodegradable scaffolds:

BIODEGRADABLE POLYMERS

Biodegradable polymers slowly degrade and then dissolve following implantation. This feature may be important for many tissue-engineering applications, since the polymer will disappear as functional tissue regenerates. For this reason, interactions of cells with a variety of biodegradable polymers have been studied.

Biodegradable polymers may provide an additional level of control over cell interactions: during polymer degradation, the surface of the polymer is constantly renewed, providing a dynamic substrate for cell attachment and growth.

[Desmond84]

Some questions that still remain to be answered are:

1) Most of their work is based on maintaining adult hair follicle (HF) Keratinocyte gene expression close to 100% with little emphasis on the gene expression of DP cells! Have they carried out gene analysis studies on DP cells or not? I'm still trying to read their paper in full but it remains elusive.

2) From what I could gather, the cells for these studies were sourced from mice whiskers not human tissue. (I may be wrong. I'll be doing more research on that) But have they actually done pre-clinical work using human DP & Keratinocyte cells?

3) Aderans went down the path of Keratinocyte/DP culturing to no avail. How different is the Taiwanese culturing method to Aderans? Do we have access to Aderans culturing methods during their last trials via patents, etc? If so, please post it here guys. Lets look into it.

__________________________________________________ _____________

Interestingly, they kind of hint at failures of Aderans and offer a solution. Designing a core-shell structure with a ball of DP cells on the inside covered by a layer of Keratinocytes on the outside:

"The present invention also confirmed observations of previous publications that, when adult keratinocytes and DP cells were simply mixed together, HFs barely grew. Therefore, the present invention provided a method for the manufacture of microtissues for inducing the growth of a hair follicle, in which keratinocytes and DP cells were cultivated simultaneously to form a core-shell structure with keratinocytes on the outer surface and DP cells in the center. Such microtissues could be transplanted in human subjects and effectively grown into HFs."

Here's more on their method of engineering a Core-shell structure:

"The method of the present invention also reveals an interesting and important feature that cells have a non-randomly compartmented distribution in the microtissues: DP cells are preferentially located in the center and keratinocytes are sorted to the surface. The spontaneously formed layered structure is similar to the natural three dimensional organization of the hair bulb: a shell of keratinocytes surrounding the core of aggregated DP cells. It has been shown that, compared to random mixture of epithelial cells and mesenchymal cells, pre-patterned compartment distribution of epithelial cells and mesenchymal cells within an organ germ in vitro can facilitate epithelial-mesenchymal interaction. In the hanging drop culture, though DP and keratinocytes are able to aggregate into compact microtissues, keratinocytes can not be efficiently sorted out to the surface and are randomly mixed up with DP cells (FIG. 5B). It has been shown that the close intercellular contact between DP cells maintained in an aggregated state s vial to the preservation of its function and HF induction ability. Additionally, the close interaction between keratinocytes and DP cells is also indispensable for the maintenance of normal growth and differentiation of HF keratinocytes. Hence, the formation of the layered structure may help to preserve the aggregated state of DP cells as well as to facilitate the epithelial-mesenchymal interaction through contact between DP and keratinocytes. Compared with other systems that employ extracellular matrix to pattern cells into folloculoid microspheres or HF germs to maintain epithelial-mesenchymal interaction for pharmacological testing, the method of the present invention can help to simply and economize the procedures for production of follculoid microtissues and other epithelial organ germs.

[Desmond84]

Here's their next mind blowing study 2 years later (in 2010):

High-throughput reconstitution of epithelial-mesenchymal interaction in folliculoid microtissues by biomaterial-facilitated self-assembly of dissociated heterotypic adult cells.

The aim of this study was to develop a method for efficient production of folliculoid keratinocyte-dermal papilla (DP) microtissues to facilitate epithelial-mesenchymal interaction.

The behavior of DP cells and adult keratinocytes from hairless skin on poly(ethylene-co-vinyl alcohol) (EVAL) surface was investigated. Keratinocytes, poorly adherent both to substrate and between homotypic cells, become suspended disperse cells after homotypic cell seeding. Seeded simultaneously, keratinocytes and DP cells are able to aggregate into spheroidal microtissues. Dynamical analysis shows that DP cells act as a carrier in the process due to the heterotypic intercellular adhesion. DP cells attach faster to EVAL and start to aggregate. Keratinocytes adhere to DP cells and are then carried by DP cells to form initial hybrid aggregates. Due to the high motility of DP cells, these hybrid aggregates move collectively as clusters and merge into larger spheroids which subsequently detach from the substratum and can be easily collected.

Compared with random cell distribution in spheroids generated in hanging drops, these hybrid spheroids have a preferential compartmented core-shell structure: an aggregated DP cell core surrounded by a keratinocyte shell. In addition to ameliorated DP signature gene expression, keratinocytes show down-regulated epidermal terminal differentiation and enhanced follicular differentiation. Functionally, these microtissues are able to grow hairs in vivo. This work sheds light on the complex effects and dynamics of cell-cell and cell-substratum interaction in the patterning of heterotypic cells into tissue forms and is of potential to be applied to mass generation of other epithelial organ primordia in vitro.

__________________________________________________ ______________

Oh my! Did you guys read the last paragraph! They are comparing their technique of culturing DP/Keratinocytes with the hanging drop method of Jahoda/Christiano, which will be conducted 3 years after the Taiwanese paper is published!

They had already tried the hanging drop method and realised that gene expression is insufficient (22%). So they went ahead with EVAL scaffolding and significantly increased gene expression!

Guys, I'm speechless. More information to continue

[Desmond84]

Here's the abstract to their first study back in 2008:

Self-assembly of dermal papilla cells into inductive spheroidal microtissues on poly(ethylene-co-vinyl alcohol) membranes for hair follicle regeneration.

Self-aggregation is key to hair follicle (HF) induction ability of dermal papilla (DP) cells and neogenesis of HF can be achieved by transplanting DP microtissues. However, there is currently lack of a suitable system that allows efficient production of DP microtissues and analysis of DP self-aggregation in vitro. We demonstrate that, at a higher seeding cell density, poly(ethylene-co-vinyl alcohol) (EVAL) membranes facilitate DP self-assembly into many compact spheroidal microtissues that are able to induce new HFs. This self-assembling process is associated with an enhanced cell movement and a declined cell-substrate adhesivity on EVAL. A compromised cell growth is also revealed on EVAL. On the contrary, a more adherent surface allows faster cell expansion but maintains DP cells in a flat morphology. Dynamically, cell migration, intercellular collision and intercellular adhesion contribute to DP microtissue formation on EVAL. Our results suggest that, for large-scale production of DP microtissues for HF regeneration, an adhesive surface is needed for quick cell expansion and a biomaterial with a lower adhesivity is required for self-aggregation. In addition, this system can be a model for investigation of DP self-aggregation in vitro.

__________________________________________________ ______________

So they were on to the aggregation issue 5 years ahead of Jahoda/Christiano and came up with a solution that long ago. How the hell they stayed under our radar is beyond me!

[kobefan234]

Those 400 people should be 400 of us here in the forums. Lord knows we've made more of an effort to warrant a treatment than many others.

yeah the experimental trial groups should be us on the forums . spot on

[Desmond84]

Sung-Jan Lin is the Clinical Trial principal Investigator

This is a patent under his name from 2013:

http://patft.uspto.gov/netacgi/nph-Parser?Sect1=PTO1&Sect2=HITOFF&d=PALL&p=1&u=%2Fnetahtml%2FPTO%2Fsrchnum.htm&r=1&f=G&l=50&s1=8492112.PN.&OS=PN/8492112&RS=PN/8492112

"Disclosed is a method for the manufacture of microtissues, comprising the steps of: providing a biomaterial substrate; simultaneously seeding a plurality of dermal papilla (DP) cells and keratinocytes on the substrate surface with a predetermined ratio and cellular density; co-culturing for a predetermined period; and carrying the keratinocytes to the substrate surface by the dermal papilla cells, aggregating and finally form a plurality of keratinocyte-dermal papilla cell microtissues, wherein the dermal papilla cells are located in a center of the microtissue and the keratinocytes are sorted to a surface of the microtissue, and the keratinocytes are adult keratinocytes. The method can help to simply and economize the procedures for production of folliculoid microtissues with high-throughput. Once microtissues are transplanted to skin of subject, hair follicles can be regenerated."

"The inventor of the present invention has previously reported that EVAL, containing both hydrophilic and hydrophobic domains, is a unique polymer that is able to enhance the self-assembly for DP cells into spheroidal microtissues that are able to induce HF morphogenesis"

Effectiveness of current treatments for severe cases of hair loss or alopecia is limited, however, the method of the present invention uses autograft cells for mass production of microtissues with inducible HF neogenesis feature in vitro, wherein intracellular contact surface is increased to facilitate induction of specific gene expressions of adult keratinocytes toward elevated efficiency of HF differentiation during process of the three dimensional keratinocyte-DP cell microtissues formation. In the aspect of production process, only small amount of DP cells and keratinocytes are required for large scale production. These cells are mass generated and then cultivated on an EVAL membrane to form microtisssues, in which an EVAL membrane is biocompatible with no harm to human health. In addition, the method of the present invention does not require precision instruments, and no complicate techniques and manufacturing processes are involved, so that production cost can be effectively cost down. More importantly, the present invention has proved that the microtissues, when transplanted to individuals (nude mice), can develop into HF and hair sheath effectively so as to be applied to HF regeneration for the treatment of hair loss and alopecia.*

OK, there's a lot of information in this patent which will take some time to go through. The first thing that stood out to me was EVAL. What the hell is EVAL? So, here's what I dug out

EVAL which is short for Ethylene vinyl alcohol is a type of plasticy polymer used in the food industry and some surgical procedures. Here's a photo of it in case you're wondering:



BTW, EVAL is a trademark name, it's industrial name seems to be EVOH. We've all actually seen EVAL before. Apparently, in the milk carton, the middle layer is EVAL which keeps the milk in the carton and stops it from spilling out. There is then a layer of laminate on top of the EVAL on the outside.

In medicine, it is used in a liquid embolic system in interventional radiology such as Onyx. Basically it is used to embolise blood vessels.

The only paper I have managed to find on EVAL being used as a scaffold in tissue engineering is the following:

Our group has developed starch-based 3D scaffolds (Gomes et al. 2003; Pavlov et al. 2004) by melt-spinning blends of starch with ethylene vinyl alcohol copolymer (EVAL), poly(ϵ-caprolactone) (PCL) and polylactide (PLA) into fibre bundles. In this methodology, fibre mesh scaffolds are produced by applying a heat treatment to bond fibre bundles (Pavlov et al. 2004). For these scaffolds, cell survival was shown to be highly dependent on scaffold porosity, which is believed to be related to the more efficient diffusion of nutrients within the scaffold (Gomes et al. 2004a).

This paper was published back in 2004. EVAL alone is actually non-porous that is why it is so widely used in the food industry to package long shelf life product. It keeps air and moisture our of the pack. So, if porosity is so vital for cell survival, how have these guys overcome this issue? That's one for a discussion.

I'll post more info as I find them. BTW, Southern your investigative talents are amazing Keep it up brother.

TBC

[Atum]

That's exactly the reason why they can't.

We're all taking different drugs already to treat our hair loss, they need people who are not taking anything (for the most part).

I'm not. The only thing i take are vitamins, amino acids, green tea, msm and gonna try CBD and manuka antiseptic creme after derma rolling.

[southern]

Effectiveness of current treatments for severe cases of hair loss or alopecia is limited, however, the method of the present invention uses autograft cells for mass production of microtissues with inducible HF neogenesis feature in vitro, wherein intracellular contact surface is increased to facilitate induction of specific gene expressions of adult keratinocytes toward elevated efficiency of HF differentiation during process of the three dimensional keratinocyte-DP cell microtissues formation. In the aspect of production process, only small amount of DP cells and keratinocytes are required for large scale production. These cells are mass generated and then cultivated on an EVAL membrane to form microtisssues, in which an EVAL membrane is biocompatible with no harm to human health. In addition, the method of the present invention does not require precision instruments, and no complicate techniques and manufacturing processes are involved, so that production cost can be effectively cost down. More importantly, the present invention has proved that the microtissues, when transplanted to individuals (nude mice), can develop into HF and hair sheath effectively so as to be applied to HF regeneration for the treatment of hair loss and alopecia.*

[Sogeking]

Well I guess it seems this is happening for real. It would be nice to see more news coverage just to be sure!

Agreed, but IMHO this is the best news I heard in the last 2 years!

[Thinning87]

Well I guess it seems this is happening for real. It would be nice to see more news coverage just to be sure!