The Next Generation of Brain Health Diagnostics

Unlocking the power of cellular saliva to detect brain injury and dementia at the point of care.

Our Breakthrough Innovations

The first disease biomarkers discovered in saliva cells

For years, the challenge in saliva diagnostics has been the low and highly variable biomarker concentration in the standard specimen — the saliva fluid.

We have fundamentally changed saliva diagnostics by showing that Saliva contains a high concentration of live cells expressing disease biomarkers.

ADVANTAGE: Saliva with cells provides >100 times higher and 7-fold less variable biomarker concentration than saliva fluid.

This breakthrough allows our BrainGuardian™ to detect the saliva-based TBI biomarker using a rapid lateral flow test — no laboratory equipment required.

Standard saliva specimen (low biomarker concentration, no rapid test) versus saliva specimen with cells (high biomarker concentration, rapid test possible)

Standard saliva fluid yields low biomarker concentration (pg/ml). Cellular saliva yields high concentration (ng–μg/ml), enabling a rapid visual test.

Cellular composition of human saliva — Hematoxylin and Eosin stain, original magnification x100

Original magnification ×100

Cellular composition of human saliva — Hematoxylin and Eosin stain, original magnification x400, showing epithelial cells (E), granulocytes (G), monocytes (M), and lymphocytes

Original magnification ×400

Cellular composition of human saliva. Hematoxylin & Eosin stain. Adult human saliva contains approximately 2 million cells/ml comprising epithelial cells (E), granulocytes (G), monocytes (M) and lymphocytes.

References

S. Southern, J Hum Virol. (1998) 1(5):328-37. Milk-borne transmission of HIV. Characterization of productively infected cells in breast milk and interactions between milk and saliva.

Southern, S. et al. (2002). Cellular Mechanism for Milk-Borne Transmission of HIV and HTLV. In: Advances in Experimental Medicine and Biology, vol 503. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-0559-4_21

Our Breakthrough Innovations

The first digital IHC assay for biomarkers in saliva cells and fluid

ADVANTAGE: high throughput, quantitative saliva biomarker analysis

standardized saliva microarray | 4 data points per slide | automated slide scan | AI-powered image analysis

Digital IHC assay of saliva with cells - four panels: a) digitized IHC slide with saliva microarray, b) Mucin1 in epithelial cells, c) segmented image, d) staining intensity map

Digital IHC assay of saliva with cells. a, Digitized IHC slide with saliva microarray (4 fields). Field 4 (green frame) was selected for image analysis. b, Mucin1 in epithelial cells (red immunostain). c, Segmented image: magenta — epithelial cells, dark blue — non-epithelial cells, cyan — saliva fluid. d, Staining intensity map: white — Mucin1 signal.

Accuracy and reliability of the saliva IHC assay - standard calibration curve over 40-fold concentration range with R-squared 0.9723 and mean CV 16 percent

Accuracy and reliability of the saliva IHC assay. Accuracy 97%, CV 16%. a, Image analysis of Muc1 signal (magnification ×200). b, Standard calibration curve for the Muc1 IHC assay shows a 40-fold linear dynamic range.

Hamamatsu NanoZoomer S210 whole-slide scanner for digital pathology Hamamatsu
Digital image of immunostained saliva — saliva microarray slide with Region of Interest profiles for the Hsp27 biomarker

Digital image of immunostained saliva microarray

Southern SO, inventor; Gaia Medical Institute, assignee. Health test for a broad spectrum of health problems. United States patent 0705097 (2020).

Our Breakthrough Innovations

The first image analysis algorithm for saliva
AI powered by Visiopharm

Visiopharm

ADVANTAGE: accurate automated measurement of biomarker signal in saliva cells and fluid

Digital image of saliva IHC stain

Digital image of a saliva IHC stain showing epithelial cells

AI APP:
100% accurate in 1 week

Visiopharm AI APP segmentation: 100% accurate classification of epithelial cells (magenta) and non-epithelial cells (dark blue)

Conventional RGB APP:
inaccurate after 3 months of effort

Conventional RGB APP segmentation: inaccurate classification of epithelial and non-epithelial cells in the saliva image

Rapid optimization of saliva image segmentation using VISIOPHARM AI APP. Visiopharm AI APP: rapid development time, 100% accurate segmentation of epithelial cells (magenta) and non-epithelial cells (dark blue) in the saliva image. Conventional RGB APP: slow development time, incorrect classification of epithelial and non-epithelial cells in the digital saliva image.

BREAKTHROUGH INNOVATIONS

Objective discovery of TBI biomarkers in saliva

The traditional way to identify a disease biomarker has been to measure of the components of the injured tissue leaking into blood and other body fluids (e.g., troponin after heart attack or GFAP/UCHL-1 after TBI).

We used our new method STRESS RESPONSE PROFILING (SRP) (US Patent 8,518,649) for a large-scale data mining of systemic response to TBI to objectively discover new TBI biomarkers in saliva.

Sarka O. Southern, W. Frank Peacock, and Ava M. Puccio. "Saliva biomarkers of traumatic brain injury." Biomarkers for Traumatic Brain Injury. Academic Press, 2020. pp. 333-353.

Iceberg diagram showing the small known part of TBI injury versus the much larger, unknown part, with TBI biomarkers in saliva highlighted in the unknown part

Stress Response Profiling (SRP) measures the systemic biological response to TBI using an array of 100 surrogate biomarkers that monitor activation of principal homeostatic pathways with known links to cellular and molecular responses to TBI. The SRP method for biomarker discovery has been successfully applied to other diseases such as dementia, showing that it is innovative in a broad sense. The table below shows the 11 systemic responses to TBI mapped to their homeostatic pathways and corresponding SRP biomarkers.

Stress Response Profiling table showing 11 systemic responses to TBI, their corresponding homeostatic pathways, and a heatmap of 100 SRP biomarkers
Our Breakthrough Innovations

Saliva IHC assay powers saliva biomarker discovery and clinical validation

Saliva biomarker for mild TBI/concussion

IHC assay of mTBI biomarker in clinical saliva samples - six panels a-f comparing a control patient (a-c) and an mTBI patient (d-f) across mTBI biomarker, negative control, and positive control

IHC assay of mTBI biomarker in clinical saliva samples. a-c, ED control. Emergency Department patient with a general complaint without head injury. d-f, mTBI. Emergency Department mTBI patient 10h post injury. a and d, mTBI biomarker. b and e, Negative control, no primary antibody. c and f, Positive control, reference saliva protein Mucin1. Original magnification ×400.

Reference

  • Southern SO, Peacock WF, Puccio AM. Chapter 23 — Saliva biomarkers of traumatic brain injury. In: Wu AHB, Peacock WF, eds. Biomarkers for Traumatic Brain Injury. Academic Press; 2020:333–353.
Our Breakthrough Innovations

Saliva IHC assay powers saliva biomarker discovery and clinical validation

Saliva biomarker for Neurocognitive Disorder due to mTBI (NCDT)

persistent cognitive, physical, and emotional impairments after mTBI

IHC assay of NCDT biomarker in clinical saliva samples - four panels: a-b healthy control with baseline biomarker expression, c-d Neurocognitive Disorder due to mTBI (NCDT) with high biomarker expression in saliva epithelial cells

IHC assay of NCDT biomarker in clinical saliva samples. a-b, Healthy control. Baseline expression of the NCDT biomarker in saliva epithelial cells (arrow). c-d, Neurocognitive Disorder due to mTBI (NCDT). Saliva epithelial cells express and secrete high level of the NCDT biomarker (arrow). Original magnifications: ×100 (a, c) and ×400 (b, d).

Reference

  • Southern SO, Peacock WF, Puccio AM. Chapter 23 — Saliva biomarkers of traumatic brain injury. In: Wu AHB, Peacock WF, eds. Biomarkers for Traumatic Brain Injury. Academic Press; 2020:333–353.
Our Breakthrough Innovations

Saliva cell culture: a “biological mirror” of the human mouth

ADVANTAGE: rapid optimization of oral film drug delivery and derisking adverse drug reactions during Phase I clinical trials

Key areas for early-stage screening of film components using saliva cell culture

  1. Optimizing In Vivo Dissolution (Saliva Interaction)
  2. Overcoming Bioavailability & Permeation Barriers
  3. Engineering Mucoadhesion
  4. Irritation Screening: drug-induced cellular stress or cytotoxicity

Example: in vitro oral drug safety test

United States patents 8518649 (2013) and 8771962 (2014)

IHC assay of saliva cellular stress - three panels: A baseline stress biomarker level, B elevated biomarker level 30 minutes after drug spike, C high cytotoxicity 4 hours after drug spike, in a primary culture of normal human saliva cells

IHC assay of saliva cellular stress. Patented Stress Response biomarker panel was measured in a primary culture of normal human saliva cells. A, Baseline stress biomarker level. B, Elevated biomarker level 30 min after drug spike. C, High level of cytotoxicity 4 hours after the drug spike. Original magnification: ×400.

Our Breakthrough Innovations

BrainGuardian™ — platform saliva LFA technology

Prototype: we have prototyped visual LFA tests for rapid quantitative detection of TBI and dementia biomarkers in human saliva to show feasibility of our commercial products.

BrainGuardian™ platform: commercial saliva LFA tests for TBI and dementia (Mild Cognitive Impairment and Alzheimer’s Disease).

BrainGuardian TBI/Concussion Test and MCI/Dementia Test kits with saliva collection swab and tube
BrainGuardian TBI test cassettes showing a positive result with two lines and a negative result with one line

Two lines means positive. One line means negative. Results you can read in minutes — just like a rapid Covid test.

Key advantages of LFA device

  • Simplicity: easy to use test kit in outpatient settings, rapid result (10–15 min) and low cost ($20–30/test)
  • Economy: inexpensive commercial development and manufacture
  • Known and achievable FDA path
  • Easy to use in point-of-care, home and field settings: patient interaction is not required to perform the test aside from saliva sample self-collection
  • Hardiness: ambient temperature storage — no refrigeration needed, multi-year shelf life
  • Wide consumer acceptability: billions of LFA test strips are consumed each year to test for Covid, pregnancy, ovulation and cardiac markers among others
THE BrainGuardian™ ADVANTAGE

The BrainGuardian™ Test for TBI

No TBI
Left Behind

Blood tests and CT scan are positive only in TBI with brain bleed.

BrainGuardian provides a solution for the “Silent Majority” with undiagnosed TBI. It is the one-stop solution for the entire TBI spectrum, from sport concussions to TBI with or without a brain bleed.

Extended Diagnostic
Window

Blood tests can detect TBI biomarkers for up to 24 hours after injury.

BrainGuardian detects saliva biomarker of TBI from 1 hour up to 9 days after injury — critical for patients with delayed-onset symptoms.

Noninvasive Saliva
Collection

Point-of-Care ready for
Any Environment

BrainGuardian uses a simple, non-invasive saliva swab for a rapid "Yes/No" answer on the sideline, in the clinic, at home or on the battlefield—no needles or specialized imaging required. No medical training required. Safe for children.

Unmatched
IP protection

9 issued US and international patents: Synergistic Ecosystem of 3 functional clusters.

Cluster A: Continuous Discovery Engine (Platform IP).

Cluster B: Diagnostic Mechanics (Method IP).

Cluster C: Clinical Products (Application IP) specific revenue streams —saliva tests for TBI and dementia.

Head-to-Head

The Only Rapid, Consumer-Ready Solution

Others require a lab, a blood draw, and equipment. We just need a saliva swab.

Feature BrainGuardian™ Abbott i-STAT GE CT Scan
Detects sport concussion Yes Not indicated Not indicated
Diagnostic window 1 hour to 9 days 12–24 hours 1–24 hours
Sample type Saliva (noninvasive) Blood draw None (radiation)
Time to result 15 minutes 15–60 min 30–60 min
Cost $30–$50 $309+ $300–$4,800
Safe for children Yes Not indicated Radiation concerns
Point-of-care setting Professional & consumer Professional only Hospital only
CLINICAL VALIDATION

Validated by the Key Opinion Leaders in TBI diagnostics

Three independent NIH-funded studies at leading U.S. academic centers have validated our saliva TBI biomarker.

94%
Accuracy across 3 NIH studies
100%
Sensitivity — no missed concussion
2,128
Prospective saliva samples

N=1,059 men and women age 8–70 (mTBI and controls) were enrolled in sport concussion clinics and Emergency Departments at the University of Pittsburgh UPMC; Children’s Hospital of Philadelphia (CHOP), University of Pennsylvania, Ben Taub Hospital and Texas Children’s Hospital, Baylor College of Medicine in Houston, University of California San Diego, and University of Arkansas.

TBI cases included sport concussion, fall, motor vehicle crash, and assault. Controls included mild traumatic orthopedic injury (broken bones, no head injury), uninjured ER patients, healthy non-athletes and athletes (uninjured players and preseason athletes) — providing strong specificity data for the FDA.

INTELLECTUAL PROPERTY PORTFOLIO

Nine issued patents protect the company’s foundational technology

United States patent certificate with ribbon
New icon
9. Health test for a broad spectrum of health problem US Patent 12360122. Issued July 15, 2025
Key IP: Saliva based diagnostic biomarkers for TBI and dementia
8. Systems and methods for analyzing persistent homeostatic perturbations US Patent 10802028, October 13, 2020
Key IP: Saliva assay of dehydration biomarkers
7. Health test for a broad spectrum of health problems US Patent 0705097, July 7, 2020
Key IP: Digital biomarker immunoassays using image analysis
6. Health test for a broad spectrum of health problems US Patent 10317416, June 11, 2019
Key IP: Biomarker analysis using saliva with cells
5. Health test for a broad spectrum of health problems European patent EP 2660 596 B1, November 30, 2018
Key IP: Methods and devices for saliva diagnostics
4. Health test for a broad spectrum of health problems US Patent 9874573, January 18, 2018
Key IP: Saliva biomarkers for dehydration, heart failure and kidney disease
3. Health test for a broad spectrum of health problems US Patent 9176149, November 3, 2015
Key IP: Methods for biomarker immunoassays using microscopy slides
2. Health test for a broad spectrum of health problems US Patent number 8771962, July 8, 2014
Key IP: Methods and devices for identifying panels of saliva biomarkers associated with a medical condition or disorder
1. Systems and methods for analyzing persistent homeostatic perturbations US Patent 8518649, August 27, 2013
Key IP: The Stress Response Profiling method for objective discovery of disease biomarkers

See Who’s Behind the Science

Our clinical pathways are guided by Key Opinion Leaders who set the standard of care for concussion in the U.S.

Meet the Team